Horiba Accهوريبا

Hematology Analyzer
Performance and Reference:

Tools for Accreditation
Ref: RAL110BEN
Performance and Reference:
Tools for Accreditation
Performance and Reference: Tools for Accreditation

Ref: RAL110BEN
Contents
Foreword..........................................................................................................................................3
1. Revisions....................................................................................................................................4
2. Legal Information....................................................................................................................5
Requirements Relating to Management...................................................................7

1. Quality Management System.............................................................................................8
2. Processing Documents........................................................................................................9
3. Review of Contracts............................................................................................................10
4. External Services and Supplies.......................................................................................11
5. Continuous Improvement..................................................................................................12
6. Quality Records and Technical Records.....................................................................13
7. Certificates..............................................................................................................................16
Technical Requirements....................................................................................................21
1. Personnel..................................................................................................................................22
2. Laboratory Premises and Environmental Conditions............................................23
3. Laboratory Equipment........................................................................................................26
4. Pre-analytical Procedures.................................................................................................57
5. Analytical Procedures.........................................................................................................58
6. Ensuring Quality of Examination Procedures...........................................................59
7. Results Report........................................................................................................................65
Appendices..................................................................................................................................67
1. Glossary of Terms.................................................................................................................68
2. References...............................................................................................................................78
Performance and Reference: Tools for Accreditation i

Ref: RAL110BEN
ii Performance and Reference: Tools for Accreditation
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Foreword
1. Revisions....................................................................................................................................4
2. Legal Information....................................................................................................................5
2.1. Declaration of Conformity...........................................................................................................5
2.2. Notice of Liability........................................................................................................................5
2.3. Trademarks.................................................................................................................................5
2.4. Graphics......................................................................................................................................5
2.5. Document Symbols.....................................................................................................................6
2.6. Copyright ® 2011 by HORIBA ABX SAS....................................................................................6
2.7. Document Intended Use.............................................................................................................6
Performance and Reference: Tools for Accreditation 3

Ref: RAL110BEN
Foreword
Revisions
1. Revisions
Index Reference Software Version Date

A RAL110AEN 1.1.X June 2010
B RAL110BEN 2.1.X March 2011
All information included in this document is current as of the date of creation of this version. Changes
that may occur will be available on www.horiba.com.
To update a paper document, please contact your local HORIBA Medical representative.
4 Performance and Reference: Tools for Accreditation

Ref: RAL110BEN
Foreword
Legal Information
2. Legal Information
2.1. Declaration of Conformity
This instrument responds to the Standards and Directives named in the Declaration of Conformity.
Latest version of the EC Declaration of Conformity for this instrument is available on www.horiba.com.
2.2. Notice of Liability
The information in this manual is distributed on an "As Is" basis, without warranty. While every
precaution has been taken in the preparation of this manual, HORIBA Medical will not assume any
liability to any persons or entities with respect to loss or damage, caused or alleged to be caused
directly or indirectly by not following the instructions contained in this manual, or by using the
computer software and hardware products described herein in a manner inconsistent with our
product labelling.
2.3. Trademarks
Linux is a registered trademark of Linus Torvalds.

Other product names mentioned within this publication may be trademarks or registered trademarks
of their respective owners.
2.4. Graphics
All graphics including screens and printouts, photographs are for illustration purposes only and are
not contractual.

Ref: RAL110BEN
Foreword
Legal Information
2.5. Document Symbols
To alert the operator of potentially hazardous conditions, symbols described in this chapter are
provided wherever necessary throughout the manual.
Emphasizes information that must be followed to avoid hazard to either the operator or the
environment, or both.
Emphasizes information that must be followed to avoid possible damage to the instrument
or erroneous test results.
Emphasizes information that can be helpful to the operator before, during or after a
specific operational function.
2.6. Copyright ® 2011 by HORIBA ABX SAS
All rights reserved. No part of this publication may be reproduced or transmitted in any form or by any
means, electronic, mechanical, photocopying, recording, or otherwise, without the prior written
permission of HORIBA Medical.
HORIBA ABX SAS

Parc Euromédecine - Rue du Caducée
B.P. 7290
34184 MONTPELLIER Cedex 4 - FRANCE
Phone: +33 (0)4 67 14 15 16
Fax: +33 (0)4 67 14 15 17
2.7. Document Intended Use
This document is intended for clinical laboratories, to help them implementing their accreditation
process and to provide them with the necessary references and reference documents based on the
standard ISO 15189 : 2007 requirements. The paragraphs of this standard specified hereafter are
associated to the response given by HORIBA Medical, in order to facilitate the accreditation process.
This document is applicable to both ABX Micros ES 60 and ABX Micros 60.
Performance data is also available in this document.

Ref: RAL110BEN
Requirements Relating to Management
1. Quality Management System.............................................................................................8
2. Processing Documents........................................................................................................9
3. Review of Contracts............................................................................................................10
4. External Services and Supplies.......................................................................................11
5. Continuous Improvement..................................................................................................12
6. Quality Records and Technical Records.....................................................................13
7. Certificates..............................................................................................................................16

Ref: RAL110BEN
Quality Management System
1. Quality Management System
■ § 4.2.5 of the standard ISO 15189 : 2007

Laboratory management shall establish and implement a programme that regularly monitors and
demonstrates proper calibration and function of instruments, reagents and analytical systems. It
shall also have a documented and recorded programme of preventive maintenance and calibration
(see 5.3.2), which, at a minimum, follows manufacturer's recommendations.
Refer to the user manual (RAB237) for the use and consultation of the records of events linked to the
instrument.

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Processing Documents
2. Processing Documents

The laboratory shall define, document and maintain procedures to control all documents and
information (from internal and external sources) that form its quality documentation. A copy of each
of these controlled documents shall be archived for later reference and the laboratory director shall
define the retention period. These controlled documents may be maintained on any appropriate
medium – including, or not, paper. National, regional and local regulations concerning document
retention could apply.
HORIBA Medical provides an electronic version or a paper version (on demand) of the user manual for
ABX Micros ES 60:
■ The reference of the English version of the ABX Micros ES 60 user manual is RAB237XEN (X being
the revision index of the document).
■ The English user manual RAB237XEN is included in the multilingual user CD-Rom
ABX Micros ES 60: RAX061X (X being the revision index of the CD-Rom).
■ The English version of the ABX Micros ES 60 daily guide is available under the reference
RAB238XEN (X being the revision index of the document).
A help is available in the instrument application ("?" icon in the toolbar).
The reference of the current version of the ABX Micros ES 60 software is given in the revision table of
the Foreword section. HORIBA Medical provides reagent leaflets and Material Safety Data Sheets for
reagents, controls and calibrators in a multilingual electronic version. These documents are available
on the CD-Rom RAX055X (X being the revision index of the CD-Rom) which is supplied with the
instrument. These documents are also available online at www.horiba.com.

Ref: RAL110BEN
Review of Contracts
3. Review of Contracts
■ § 4.4 of the standard ISO 15189 : 2007

4.4.1 Where a laboratory enters into a contract to provide medical laboratory services, it shall
establish and maintain procedures for review of contracts. The policies and procedures for these
reviews leading to a change in the arrangements for examinations or contracts shall ensure that:
■ a) requirements, including the methods to be used, are adequately defined, documented and
understood (see 5.5);
■ b) the laboratory has the capability and resources to meet the requirements;
■ c) appropriate procedures selected are able to meet the contract requirements and clinical
needs (see 5.5).
In reference to b), the review of capability should establish that the laboratory possesses the
necessary physical, personnel and information resources, and that the laboratory's personnel have
the skills and expertise necessary, for the performance of the examinations in question. The review
may also encompass results of earlier participation in external quality assurance schemes using
samples of known value in order to determine uncertainties of measurement, limits of detection,
confidence limits, etc.
4.4.2 Records of reviews, including any significant changes and pertinent discussions, shall be
maintained (see 4.13.3).
All contracts (maintenance, remote assistance policy, suppliers charter) signed with HORIBA Medical
must be retained.

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External Services and Supplies
4. External Services and Supplies

Laboratory management shall define and document its policies and procedures for the selection
and use of purchased external services, equipment and consumable supplies that affect the quality
of its service. Purchased items shall consistently meet the laboratory's quality requirements.
National, regional or local regulations may require records of purchased items. There shall be
procedures and criteria for inspection, acceptance/rejection and storage of consumable materials.
ABX Micros ES 60, its reagents, controls and calibrators comply with the standards and directives
that are included in each declaration of conformity.
Latest version of the CE Declaration of Conformity for this instrument is available online at
www.horiba.com.
The quality management system of HORIBA Medical is certified with respect to the standards
ISO 13485 : 2003 and ISO 9001 : 2008.

There shall be an inventory control system for supplies. Appropriate quality records of external
services, supplies and purchased products shall be established and maintained for a period of
time, as defined in the quality management system. This system shall include:
■ a) the recording of lot numbers of all relevant reagents, control materials and calibrators,
■ b) the date of receipt in the laboratory and the date the material is placed in service.
These documents must be kept for 12 to 18 months depending on the country and the frequency
of audits.

The laboratory shall evaluate suppliers of critical reagents, supplies and services that affect the
quality of examinations and shall maintain records of these evaluations and list those approved.

Ref: RAL110BEN
Continuous Improvement
5. Continuous Improvement

Laboratory management shall provide access to suitable educational and training opportunities for
all laboratory personnel and relevant users of laboratory services.
To meet this requirement, HORIBA Medical has a training center that enables laboratory staff to
receive appropriate training for the correct use of the machines. This structure makes it possible to
undergo user training, on the client’s premises; these training courses are carried out at the time of
installation. Also, complete training courses (operation, result interpretation, maintenance) are
available in the HORIBA Medical training centers. All of these training courses have specific programs,
documentation, and registrations.

Ref: RAL110BEN
Quality Records and Technical Records
6. Quality Records and Technical Records

The laboratory shall have a policy that defines the length of time various records pertaining to the
quality management system and examination results are to be retained. Retention time shall be
defined by the nature of the examination or specifically for each record.
National, regional and local regulations may apply.
These records may include but are not limited to the following:
■ a- request forms (including the patient chart or medical record only if used as the request form);
■ b- examination results and reports;
■ c- instrument printouts;
■ d- examination procedures;
■ e- elaboratory work-books or sheets;
■ f- accession records;
■ g- calibration functions and conversion factors;
■ h- quality control records;
■ i- complaints and action taken;
■ j- records of internal and external audits;
■ k- external quality assessment records/interlaboratory comparisons;
■ l- quality improvement records;
■ m- instrument maintenance records, including internal and external calibration records;
■ n- lot documentation, certificates of supplies, package inserts;
■ o- incident/accident records and action taken;
■ p- staff training and competency records.
§ c, g, h, m: Refer to the ABX Micros ES 60 user manual: in Setup section for the printing conditions,
and in Maintenance and Troubleshooting section for the log entries printouts (calibration, reagents,
etc).
§ k: HORIBA Medical proposes a Quality Control Program for the inter-laboratory comparison of Daily
Internal Control results via its website.
§ n: HORIBA Medical Latest versions of these documents are available online at www.horiba.com.
§ p: Following each of the training courses undertaken in accordance with the quality control
procedures of HORIBA Medical, a certificate of attendance and a certificate of training will be
awarded.

Management review shall take account of, but not be limited to:
■ a- follow-up of previous management reviews;
■ b- status of corrective actions taken and required preventive action;
■ c- reports from managerial and supervisory personnel;
■ d- the outcome of recent internal audits;
■ e- assessment by external bodies;
■ f- the outcome of external quality assessment and other forms of interlaboratory comparison;
■ g- any changes in the volume and type of work undertaken;
■ h- feedback, including complaints and other relevant factors, from clinicians, patients and other
parties;
■ i- quality indicators for monitoring the laboratory’s contribution to patient care;

Ref: RAL110BEN
■ j- nonconformities;
■ k- monitoring of turnaround time;
■ l- results of continuous improvement processes;
■ m- evaluation of suppliers.
Shorter intervals between reviews should be adopted when a quality management system is being
established. This will allow early action to be taken in response to those areas identified as requiring
amendment of the quality management system or other practices.
Identification of the Company

Company name: HORIBA ABX SAS Subsidiary of HORIBA Group
Date of creation: Sept 30th, 1983 (Kyoto, Japan)
Address:
HORIBA Medical
Parc Euromédecine - Rue du Caducée
B.P. 7290
34184 MONTPELLIER Cedex 4 - FRANCE
Phone: +33 (0)4 67 14 15 16
Fax: +33 (0)4 67 14 15 17
Website: http://www.horiba.com/medical/
Legal type of company: SAS (Société par Actions Simplifiée)

APE/NAF Code: 2651B
Trade register: RCS Montpellier 328 031 042
Training Center agreement N°: N/A
Company Managers
President Bertrand de Castelnau
email: katia.krouk@horiba.com
Tel. +33 (0)4 67 14 15 38 (secretariat)
International Marketing Director Alfredo Braga

email: alfredo.braga@horiba.com
Tel. +33 (0)4 67 14 16 86 (secretariat)
International Technical Service Manager Jean-Louis Taravella

email: jean-louis.taravella@horiba.com
Tel. +33 (0)4 67 14 15 63
Quality & Regulatory Affairs Director Arnaud Pradel

email: arnaud.pradel@horiba.com
Tel. +33 (0)4 67 14 17 24
Reacto-Vigilance Manager Arnaud Pradel

email: arnaud.pradel@horiba.com
Tel. +33 (0)4 67 14 17 24
Main Activities sectors

Clinical analysis devices aimed at laboratories of medical analysis.
Medical devices of in vitro diagnostics.

Ref: RAL110BEN
Policy related to Quality

HORIBA ABX SAS is SGS certified as referenced below:
■ ISO 9001 - 2008 edition
■ ISO13485 - 2003 edition
■ ISO14001 - 2004 edition
■ OHSAS 18001 - 2004 edition
Refer to the Certificates chapter hereafter.
Reagents/Consumables Information
The reagents are listed in an annual catalogue available in paper format and on the website.
The reagents are all CE marked in conformity with the 98/79/CE directive.
HORIBA Medical provides the reagents notices and the security information sheets of the reagents
and blood controls and calibrators in a multi-language electronic version.
These documents are available on the CD-Rom provided with the analyser.
These documents are also available on www.horiba.com

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Certificates
7. Certificates

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Certificates

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Certificates

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Certificates

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Certificates

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Technical Requirements
1. Personnel..................................................................................................................................22
2. Laboratory Premises and Environmental Conditions............................................23

2.1. Operating Conditions................................................................................................................23
2.2. Storage Conditions for the Reagents........................................................................................23
2.3. Environmental protection..........................................................................................................24
3. Laboratory Equipment........................................................................................................26
3.1. Calibration.................................................................................................................................26
3.2. Maintenance..............................................................................................................................26
3.3. Energy Consumption................................................................................................................26
3.4. Waste Disposal.........................................................................................................................27
3.5. Performance..............................................................................................................................27
3.6. Labeling.....................................................................................................................................54
3.7. Individual Machine Data Sheets................................................................................................55
3.8. Manuals and Instructions for Use.............................................................................................55
3.9. Disposal of Chemical Products and Biological Materials.........................................................56
3.10. Use of Computers for the Analysis Data.................................................................................56
4. Pre-analytical Procedures.................................................................................................57
4.1. Specimen for Sampling.............................................................................................................57
5. Analytical Procedures.........................................................................................................58
6. Ensuring Quality of Examination Procedures...........................................................59

6.1. Traceability Chain for WBC and RBC Count............................................................................61
6.2. Traceability Chain for HGB Measurement................................................................................62
6.3. Traceability Chain for HCT Measurement.................................................................................63
6.4. Traceability Chain for PLT Count..............................................................................................64
7. Results Report........................................................................................................................65

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Personnel
1. Personnel
■ § 5.1.2.d of the standard ISO 15189 : 2007

Laboratory management shall maintain records of the relevant educational and professional
qualifications, training and experience, and competence of all personnel. This information shall be
readily available to relevant personnel, and may include:
■ a- certification or license, if required;
■ b- references from previous employment;
■ c- job descriptions;
■ d- records of continuing education and achievements;
■ e- competency evaluations;
■ f- provision for untoward incident or accident reports.
Other records available to authorized persons relating to personnel health may include records of
exposure to occupational hazards and records of immunization status.
To meet this requirement, HORIBA Medical has a training center that enables laboratory staff to
receive appropriate training for the correct use of the machines. This structure makes it possible to
undergo user training, on the client’s premises; these training courses are carried out at the time of
installation. Also, complete training courses (operation, result interpretation, maintenance) are
available in the HORIBA Medical training centers. All of these training courses have specific programs,
documentation, and registrations.

Ref: RAL110BEN
Laboratory Premises and Environmental Conditions
2. Laboratory Premises and Environmental

Conditions

The laboratory design and environment shall be suitable for the tasks carried out therein. The
environment in which the primary sample collection or examinations or both are undertaken shall
not, invalidate the results, or adversely affect the required quality, of any measurement. Laboratory
facilities for examination should permit correct performance of examinations. These include, but
are not limited to, energy sources, lighting, ventilation, water, waste and refuse disposal, and
environmental conditions. The laboratory should have procedures for checking that the
environment does not adversely affect the performance of specimen collection and equipment.
2.1. Operating Conditions

The laboratory shall monitor, control and record environmental conditions, as required by relevant
specifications or where they may influence the quality of the results. Attention should be paid to
sterility, dust, electromagnetic interference, radiation, humidity, electrical supply, temperature and
sound and vibration levels, as appropriate to the technical activities concerned.
Refer to the Introduction > Operational Conditions chapter of the user manual (RAB237) for the
conditions of use of ABX Micros ES 60.
2.2. Storage Conditions for the Reagents

Relevant storage space and conditions shall be provided to ensure the continuing integrity of
samples, slides, histology blocks, retained micro-organisms, documents, files, manuals,
equipment, reagents, laboratory supplies, records and results.
Refer to the reagent leaflets and Material Safety Data Sheets for the conditions of storage, use and
disposal. Latest versions of these reagents documents are available online at www.horiba.com.

Ref: RAL110BEN
2.3. Environmental protection
■ § 5.2.10 of ISO standard 15189: 2007

Work areas shall be clean and well maintained. Storage and disposal of dangerous materials shall
be those specified by relevant regulations. Measures shall be taken to ensure good housekeeping
in the laboratory. Special procedures and training for personnel could be necessary to that end.
Recommendations for Treatment of Liquid Effluents with Risks of Infection Produced

by Automated Systems
Biological risk
■ Bleach:
Bleach is essentially marketed in 2 forms:
■ concentrated bleach in 250 mL sachets: 9.6% active chlorine
■ ready-to-use bleach in containers: 2.6% active chlorine.
Bottles or containers of ready-to-use 2.6% active chlorine solution keep for at least one year after
opening.
Sachets of 9.6% active chlorine solution have limited stability over time and must be diluted within 2.5
to 3 months following the manufacturing date shown on the packaging.
■ Concentration and contact time: (Use of diluted bleach)
0.5% of active chlorine inactivates CTAs (Conventional Transmissible Agents) through contact of
at least 15 minutes (dilution of 1/5 using a 2.6% concentration or dilution of 1/20 using a 9.6%
concentration).
In practice, allow for overnight contact time so that the chlorine ions are able to disperse upon
contact with proteins in the treated environment.
Chemical risk
Decree 2002-540 of the environmental code (Decree no. 2007-1467).
Chemical risks must be evaluated based on laboratory activities, the types of instruments and
reagents used and the frequency of tests.
Consult safety data sheets for the reagents used to identify dangerous substances and calculate the
final concentrations of those substances for comparison with the limits set in the environmental code
(Art R 541-7, Art R 541-8 and appendices 1 and 2, Art R 541-9, Art R 541-10).
Conclusion: following elimination of biological risk and in the absence of chemical risk,
the effluent may be emptied into the sink.
In the event of residual chemical risk, the user must take the necessary measures to
dispose of this effluent
Reference:
■ Circular DGS/SD5C/DHOS/E2/DRT/CT1/CT2 no. 2004-382 dated July 30, 2004, regarding
precautions to be taken in anatomy and cytology departments, autopsy rooms, mortuary facilities
and "NCTA specialist" biological laboratories concerning the risk of transmission of conventional
transmissible agents (CTA) and non-conventional transmissible agents (NCTA).
■ Circular DGS/5C/DHOS/E2 no. 2001-138 dated March 14, 2001, regarding precautions to be
taken during treatment aimed at reducing the risks of transmission of non-conventional
transmissible agents.
■ Laboratory Biosafety Manual, Second Edition (revised), World Heath Organization. Geneva, 2003.
■ Notice from the SFHH (Société Française d'Hygiène Hospitalière – The French Society for Hospital
Hygiene) regarding the use of bleach in healthcare establishments. June 2006.

Ref: RAL110BEN
■ SFHH: Prevention of risk of infection in medical biological analysis laboratories, 2007.

■ INRS (Institut national de recherche et de sécurité – French occupational health and safety
institute) Guide « Déchets infectieux : Elimination des DASRI et assimilés, prévention et
réglementation » 2nd edition. August 2006.
■ Site Chambre syndicale nationale de l’eau de Javel (CENEJ).
■ Recommendations for Treatment of Liquid Effluents with Risks of Infection Produced by
Automated Systems. Syndicat de l’Industrie du Diagnostic (In Vitro Diagnostic Industry Union)
(SFRL). January 2010.

Ref: RAL110BEN
Laboratory Equipment
3. Laboratory Equipment

The laboratory shall be furnished with all items of equipment required for the provision of services
(including primary sample collection, and sample preparation and processing, examination and
storage). In those cases where the laboratory needs to use equipment outside its permanent
control, laboratory management shall ensure that the requirements of this International Standard
are met. When selecting equipment, account should be taken of the use of energy and future
disposal (care of the environment).
3.1. Calibration
The controls and calibrators associated with the ABX Micros ES 60, supplied by HORIBA Medical,
comply with the requirements of the standard ISO 17511 : 2003, relating to the metrological
traceability of values attributed to calibration agents and control equipment (in vitro diagnostic
medical devices) for the measurement of quantities in biological samples. This makes it possible to
guarantee that the results obtained by the ABX Micros ES 60, duly controlled and calibrated, are
sufficiently exact to enable a clinically correct interpretation and comparability over time and space.
3.2. Maintenance
Refer to Quality Management System chapter of this present document.

As a function of the actions undertaken and described in the maintenance or servicing procedures,
the HORIBA Medical technician may, by request of the Laboratory Director, return the original
documents that enabled the validation of the instrument (repeatability tests, correlation tests,
calibration, and controls).
The technician will leave the original source documents for qualification (or requalification) and
calibration in the laboratory in accordance with current procedures (installation, every four months
maintenance, yearly maintenance, check-up after intervention).
3.3. Energy Consumption
Refer to Specifications > Technical Specifications chapter of the user manual (RAB237).

Ref: RAL110BEN
3.4. Waste Disposal
For waste disposal, refer to Specifications > Waste Handling Procedure and to Introduction >
Environmental Protection chapters of the user manual (RAB237).
3.5. Performance

Equipment shall be shown (upon installation and in routine use) to be capable of achieving the
performance required and shall comply with specifications relevant to the examinations concerned.
Laboratory management shall establish a programme that regularly monitors and demonstrates
proper calibration and function of instruments, reagents and analytical systems. It shall also have a
documented and recorded programme of preventive maintenance (see 4.2.5), which, at a
minimum, follows the manufacturer's recommendations.
When manufacturers' instructions, operators' manuals or other documentation are available, they
may be used to establish requirements, for compliance with relevant standards or to specify
requirements for periodic calibration, as appropriate, to fulfil part or all of this requirement.
3.5.1. Parameters
The techniques used for counting, measuring, and detecting the various parameters are explained in
the Description and Technology section of the user manual (RAB237).
CBC Parameters Definition

WBC White Blood Cells
LYM # Lymphocytes absolute value
LYM % Lymphocytes percentage
MON # Monocytes absolute value
MON % Monocytes percentage
GRA # Granulocytes absolute value
GRA % Granulocytes percentage
RBC Red Blood Cells
HGB Hemoglobin Concentration
HCT Hematocrit
MCV Mean Corpuscular Volume
MCH Mean Corpuscular Hemoglobin
MCHC Mean Corpuscular Hemoglobin Concentration
RDW Red Distribution Width
PLT Platelets
PDW * Platelets Distribution Width
PCT * Plateletcrit
MPV Mean Platelet Volume

Ref: RAL110BEN
* PDW and PCT have not been established as indications for use in United States for this
instrument. Their use should be restricted to Research Use Only (RUO). Not for use in
diagnostic procedure.
3.5.2. Precision (Reproducibility)
Assessment Protocol for reproducibility (within-device precision)
The CLSI EP15-A2 document recommends that:

■ The assessment is performed on at least two levels, as precision can differ over the analytical
range of an assay.
■ Each level is run with three replicates over five days.
■ The order of analysis of test materials and QC for each run or day should be changed.
■ To simulate actual operation, include at least ten patient samples in each run.
The reader is referred to the CLSI EP15-A2 documents for details.
When undertaking the assessment, the data must be assessed for outliers, which are considered to
be present if the absolute difference between replicates exceeds 5.5 times the SD determined in the
preliminary precision test. If an outlier is found the pair should be rejected and the cause investigated
and resolved before repeating the run. The figure of 5.5 is derived from the upper 99.9% value of the
normalised range for differences between two populations.
Acceptable CV limits established on control blood (ABX Minotrol 16):
Parameters Low level Normal level High level

WBC < 7% < 5% < 4%
RBC < 4% < 3% < 3%
HGB < 5% < 4% < 3%
HCT < 5% < 4.5% < 4%
MCV < 4% < 3% < 2.5%
PLT < 15% < 10% < 7%
LYM% < 8% < 8% < 8%
MON% < 15% < 14% < 11%
GRA% < 12% < 4% < 3%
Mean CV obtained with several batches of control blood (ABX Minotrol 16):
These results are based on the study on precision by replicate runs of control bloods, according to
the protocol detailed in the CLSI EP15-A2 document.
ABX Micros ES 60 OT

WBC 3.29% 1.76% 1.55%
RBC 1.75% 1.25% 1.38%
HGB 1.53% 0.98% 0.8%
HCT 1.65% 1.26% 1.35%
MCV 0.69% 0.7% 0.61%
PLT 6.98% 3.38% 2.9%
LYM% 2.01% 1.84% 1.8%
MON% 6.77% 4% 4.34%

Ref: RAL110BEN

GRA% 3.7% 1.01% 0.34%
ABX Micros ES 60 CT

WBC 3.05% 1.69% 1.4%
RBC 2.04% 1.37% 1.29%
HGB 2.52% 1.43% 1.27%
HCT 1.99% 1.33% 1.33%
MCV 0.79% 0.82% 0.72%
PLT 5.95% 3.47% 2.45%
LYM% 7.29% 4.44% 5.02%
MON% 2.02% 1.92% 2.11%
GRA% 3.7% 1.06% 0.38%
Mean CV obtained with human blood

As the white blood cell count of the control bloods was different from that of human blood, a specific
CV was calculated by running human bloods ten times, the same day, under comparable and
reproducible conditions. The resulting coefficients of variation were as follows:
Parameters Level CV
LYM% 24.74% 5.86%
MON% 6.35% 10.12%
GRA% 68.13% 4.09%
3.5.3. Precision (Repeatability)
Assessment Protocol for repeatability (within-run precision)

The CLSI H26-P2 document (§5.9) recommends that:
This imprecision study is performed using apparently normal fresh whole blood whose values do not
exceed the normal reference interval for the laboratory. A sample size of 31 consecutive aspirations of
the same blood collection tube is used; smaller sample sizes produce less desirable wider confidence
intervals. At least four imprecision studies are performed on each of at least three Test Automated
Analyzers (TAA), for a total of at least 12 runs.
Repeatability:
Mean CV obtained Mean CV obtained

Parameters Specifications Level of analysis ABX Micros ES 60 ABX Micros ES 60
OT CT
WBC < 2.5 % 3.6 - 9.8x 103/mm3 1.48 % 1.65 %
RBC <2% 3.54 - 5.2 x 106/mm3 0.96 % 1.19 %
HGB < 1.5 % 10.7 - 16 g/dL 0.51 % 1%
HCT <2% 33.6 - 46.8 % 1.07 % 1.38 %
MCV < 1.5 % 86 - 98.5 fL 0.63 % 0.57 %
PLT <5% 160 - 365 x 103/mm3 3.2 % 3.23 %
LYM% <5% 16.5 - 45 % 2.23 % 2.64 %
MON% < 10 % 4.3 - 10 % 4.67 % 7.54 %

Ref: RAL110BEN
Mean CV obtained Mean CV obtained

Parameters Specifications Level of analysis ABX Micros ES 60 ABX Micros ES 60
OT CT
GRA% <3% 46 - 80 % 1.37 % 1.42 %
3.5.4. Approach to Trueness
Trueness is defined as ‘the closeness of agreement between the average value obtained from a large
series of test results and an accepted reference value’ (ISO 3534- 1-3.12) 1 . Accuracy, on the other
hand, is defined as the ‘closeness of the agreement between the result of a measurement and a true
value of the measurand’ (VIM 93-3.5 ) 2 .
In ISO/CD 17511 3 , the concept of ‘accuracy of measurement’ relates both to trueness of
measurement and precision of measurement. The IVD-Directive 4 , on the other hand, refers to
specific analytical performance characteristics and uses the term 'accuracy' and 'trueness'
synonymously. Büttner 5 (see figure below) has presented a classification of analytical performance
characteristics and techniques for their evaluation. There is an important difference between
‘trueness’ and ‘accuracy’.
The definition of ‘uncertainty of measurement’ from the same source (VIM, 1993)2 is also widely
defined as a ‘parameter, associated with the result of a measurement, that characterises the
dispersion of the values that could reasonably be attributed to the measurand’. All laboratory tests are
subject to uncertainties inherent in the test or errors arising during performance.
Performance Kind of Term of

Characteristics Error Expression
___________ TRUENESS ___________ SYSTEMATIC ___________ BIAS

___________
ERROR
___________ ___________ TOTAL ___________ UNCERTAINTY

ACCURACY ERROR
___________ PRECISION ___________ RANDOM ___________ STANDARD

ERROR DEVIATION
Expression of analytical specific performance characteristics (from Büttner, 1994)
Bias calculation
Trueness, quantified by the bias, was calculated by comparing the mean obtained (m) in
reproducibility with respect to the expected target value (V), assimilated to the true value of the tested
sample. For the white blood cell differential count, the bias was calculated with respect to the value
obtained using a manual count on 200 cells.
1 Statistics-Vocabulary and symbols-Part 1: Probability and general statistical terms. ISO 3534-1, Geneva: International
Organization for Standardization (ISO), 1993.

2 International Vocabulary of Basic and General Terms in Metrology (VIM), 2nd edition. Geneva: International Organization for
Standardization (ISO), 1993.

3 In Vitro Medical Devices-Measurement of Quantities of Biological Origin-Metrological Traceability of Values Assigned to
calibrators and Control Materials, ISO/CD 17511, Geneva: International Organization for Standardization (ISO), 2000.
4 Directive 98/79/EC of the European Parliament and of the Council of 27 October 1998 on in vitro diagnostic medical devices
(Official Journal of the European Communities L 331, 7.12.98).

5 Büttner J: Reference Materials and Reference Methods in Laboratory Medicine: A Challenge to International Cooperation, Eur J
Clin Chem Clin Biochem, 32,8: 571-577, 1994.

Ref: RAL110BEN
The bias was calculated using data obtained on a selection of analyzers. The indicated bias limit is
calculated from the control charts to two standard deviations.
(m - V)
Bias in % = ______ x 100
V
Bias on the blood counts undertaken with several batches of ABX Minotrol 16:
CBC Parameters Low level Normal level High level

Level (103/mm3) 2 7.5 20
WBC Bias limit < 9% < 4% < 4%
Bias obtained 2.64% -0.74% 0.48%
Level (106/mm3) 2.4 4.6 5.7
RBC Bias limit < 6% < 3% < 4%
Bias obtained 1.45% 0.84% 0.95%
Level (g/dL) 6.3 13.5 18.2
HGB Bias limit < 6% < 3% < 3%
Bias obtained 1.77% -0.14% 0.32%
Level (%) 17 37 50
HCT Bias limit < 8% < 4% < 5%
Bias obtained 1.56% 0.39% 1.84%
Level (fL) 71 80 90
MCV Bias limit < 5% < 3% < 4%
Bias obtained 0.2% -0.46% 0.84%
Level (%) 12.9 12.2 11.9
RDW Bias limit < 5% < 5% < 4%
Bias obtained -0.9% -2.8% -8.61%
Level (103/mm3) 80 240 480
PLT Bias limit < 8% < 8% < 9%
Bias obtained 4.84% 1.42% 3.07%
Level (µm3) 8.3 8 7.9
MPV Bias limit < 6% < 6% < 4%
Bias obtained -1.1% 1.74% 1.04%
ABX Micros ES 60 OT

Level 8 26 55
LYM% Bias limit < 25% < 20% < 30%
Bias obtained 15.2% -10.3% -16.3%
Level 2.5 5 14
MON% Bias limit < 140% < 20% < 70%
Bias obtained 126.4% 16.04% -54.9%
Level 35 65 88
GRA% Bias limit < 30% < 10% < 5%
Bias obtained 19.74% 7.6% 3.91%

Ref: RAL110BEN
ABX Micros ES 60 CT

Level 8 26 55
LYM% Bias limit < 25% < 20% < 30%
Bias obtained 11.8% -10.8% -16%
Level 2.5 5.66 14
MON% Bias limit < 140% < 20% < 70%
Bias obtained 106.8% 8.14% -55.3%
Level 38.85 65 89.75
GRA% Bias limit < 30% < 10% < 5%
Bias obtained 21.6% 8.2% 4.09%
QCP: A new service to improve your Quality Control System

HORIBA Medical is offering a new service to Clinical Laboratories: The ABX Quality Control Program.
The ABX Quality Control Program or QCP is an internet-based Inter-laboratory Comparison Program
dedicated to all of the Hematology and Biochemistry analyzers in the HORIBA Medical range. It
comprises a software program that can be accessed via the internet that enables registered
laboratories to assess the Exactitude and Precision of their analyzers. By submitting their internal
quality control results to the website of HORIBA Medical at www.horiba.com, member laboratories
can access, in real time, statistical reports comparing these results with those of their peer group. The
ABX Quality Control Program was designed to provide effective support to registered laboratories in
their quality control system and to enable them to meet the regulatory requirements that encourage
participation in inter-laboratory comparison campaigns (ISO/CEI 43-1 guide) in order to demonstrate
the analytical performance of their analyzers.
Calculation of uncertainty
The uncertainty on blood results is calculated from two components:
■ The component U1 due to the reliability approached by the reproducibility standard deviation. This
component is obtained from the values obtained with control bloods with the exception of the
differential white blood cell count for which the calculation was undertaken on human blood.
U1 = S repro
■ The component of uncertainty due to trueness, U2
Uncertainty #
U2 = _____________
√3
Uncertainty # = (m - V) : where m is the mean obtained, and V is the expected target value.
■ The combined uncertainty Uc is calculated using the following formula:
_____
Uc = √ U12 +U22
■ Expanded uncertainty = 2 * Uc

Ref: RAL110BEN
Uncertainty calculated on the blood count:

Level (103/mm3) 2 7.5 20
WBC Uncertainty # 0.16 0.35 0.71
Uc% 8.19% 4.64% 3.53%
Level (106/mm3) 2.4 4.6 5.7
RBC Uncertainty # 0.1 0.17 0.19
Uc% 3.99% 3.74% 3.39%
Level (g/dL) 6.3 13.5 18.2
HGB Uncertainty # 0.34 0.55 0.64
Uc% 5.42% 4.11% 3.51%
Level (%) 17 37 50
HCT Uncertainty # 0.86 1.66 2.23
Uc% 5.05% 4.47% 4.46%
Level (fL) 71 80 90
MCV Uncertainty # 2.02 1.75 1.86
Uc% 2.84% 2.19% 2.07%
Level (%) 12.9 12.2 11.9
RDW Uncertainty # 0.5 0.59 1.27
Uc% 3.87% 4.85% 10.64%
Level (103/mm3) 80 240 480
PLT Uncertainty # 16.06 22.82 31.57
Uc% 20.07% 9.51% 6.58%
Level (µm3) 8.3 8 7.9
MPV Uncertainty # 0.6 0.4 0.3
Uc% 7.19% 5.04% 3.86%
Uncertainty calculated on the differential white blood cell count:

Level 13.5 31.39 49.5
LYM% Uncertainty # 2.67 5.53 11.98
Uc% 19.77% 17.62% 24.21%
Level 3 7.45 14
MON% Uncertainty # 4.43 1.51 7.33
Uc% 147.72% 20.31% 52.38%
Level 39.5 61.16 80.5
GRA% Uncertainty # 9.08 6.59 6.61
Uc% 22.99% 10.77% 8.21%

Ref: RAL110BEN
3.5.5. Assessment of the Performance using the Sigma Calculation

(according to Westgard, 2001)
The capability of an analytical system is the measurement of the relationship between its true
performance and the requested performance. In the “Six Sigma” approach, the capability indicator
represents the difference between the requested performance (total allowable error TEa) and the
mean expressed in numbers of standard deviations, hence the formula:
(TEa - bias %)
Sigma = _______
CV
■ CV: Coefficient of Variation
■ TEa: total allowable error is that given by Ricos & al. 6 for a risk of 5 %
Capability calculated on the blood count:
CBC parameters TEa Ricos & al. Level Bias % CV % Sigma

2 2.64 % 3.75 % 3.19
WBC
14.6 % 7.5 0.74 % 2.31 % 6.00
(103/mm3)
20 0.48 % 1.77 % 7.98
2.4 1.45 % 1.89 % 1.56
RBC
4.4 % 4.6 0.84 % 1.79 % 1.99
(106/mm3)
5.7 0.95 % 1.61 % 2.14
6.3 1.77 % 2.63 % 0.88
HGB
4.1 % 13.5 0.14 % 2.04 % 1.94
(g/dL)
18.2 0.32 % 1.75 % 2.16
17 1.56 % 2.37 % 1.07
HCT
4.1 % 37 0.39 % 2.18 % 1.70
(%)
50 1.84 % 1.92 % 1.18
71 0.20 % 1.37 % 1.53
MCV
2.3 % 80 0.46 % 1.04 % 1.77
(fL)
90 0.84 % 0.92 % 1.60
12.9 0.90 % 1.82 % 2.04
RDW
4.6 % 12.2 2.80 % 1.72 % 1.05
(%)
11.9 8.61 % 1.86 % -2.16
80 4.84 % 9.26 % 0.92
PLT
13.4 % 240 1.42 % 4.42 % 2.71
(103/mm3)
480 3.07 % 2.77 % 3.72
8.3 1.10 % 3.56 % 1.32
MPV
5.8 % 8 1.74 % 2.49 % 1.63
(fL)
7.9 1.04 % 2.24 % 2.13
6 DESIRABLE SPECIFICATIONS FOR TOTAL ERROR, IMPRECISION, AND BIAS, DERIVED FROM BIOLOGIC VARIATION by Ricos
C, Alvarez V, Cava F, Garcia-Lario JV, Hernandez A, Jimenez CV, Minchinela J, Perich C, Simon M. "Current databases on biologic
variation: pros, cons, and progress." Scand J Clin Lab Invest 1999;59:491-500 (revised in 2008).

Ref: RAL110BEN
Graphs of capability
Capability calculated on the white blood cells:
DIF parameters TEa Ricos & al. Level Bias % Uncertainty % Sigma
14 13.8 % 2.34 % 3.68
LYM% 22.4 % 31 11.4 % 1.93 % 5.70
50 18.3 % 2.51 % 1.62
3.0 126.7 % 7.39 % -14.97
MON% 16% 7.5 1.4 % 5.42 % 2.69
14.0 41.8 % 4.47 % -5.78

Ref: RAL110BEN
DIF parameters TEa Ricos & al. Level Bias % Uncertainty % Sigma
39.5 18.6 % 4.72 % 1.97
GRA% 27.9 % 61.2 6.1 % 1.37 % 15.98
80.5 0.6 % 0.62 % 43.83
3.5.6. Specificity of the Parameters
Refer to the Specifications section for parameters definitions and to the Description and Technology
section of the user manual (RAB237) for the measurement techniques and the specificities of the
parameters of ABX Micros ES 60.
3.5.7. Field of Analysis
The device composed of the ABX Micros ES 60 is a system enabling the in vitro diagnostic analysis of
whole blood samples. Refer to the Specifications > Limitations chapter of the user manual (RAB237)
for the types of samples authorized for the ABX Micros ES 60.
3.5.8. Analytical Sensitivity
Detection limit (XP T 90-210)

It is the smallest quantity of an analyte to be examined in a sample that can be detected and
considered as being different from the value of the blank (with a given probability), but not necessarily
quantified. Two risks need to be taken into account:
■ the risk of considering the substance present in the sample when in fact its quantity is nil.
■ the risk of considering a substance absent when in fact its quantity is not nil.
Parameter Detection Limit

WBC 0.1 x 103/mm3
RBC 0.01 x 106/mm3
HGB 0.5 g/dL
HCT 0.1 %
PLT 3.5 x 103/mm3
Quantitation limit (XP T 90-210)

It is the smallest quantity of an analyte to be analyzed in a sample that can be determined
quantitatively under the experimental conditions described in the method with a defined variability
(determined coefficient of variation).
Each dilution was run ten times in a row to assess the coefficient of variation at this level of value.
Parameter CV limits Quantitation Limit

WBC 10.9 % 0.2 x 103/mm3
RBC 3.2 % 0.3 x 106/mm3
HGB 2.8 % 2 g/dL
HCT 2.8 % 5%
PLT 9.1 % 40 x 103/mm3

Ref: RAL110BEN
3.5.9. Analytical Measuring Interval (Linearity)
Linearity is the ability of an analytical process to provide a measurement proportional to the

measurand (analyte) being quantified over a defined range of concentrations. Strictly speaking, the
concept of linearity is applicable only to HB measurements. Statisticians have pointed out that "Some
of the statistical assumptions for estimating and testing in linear regression are violated in analytical
methods that count particles per unit of volume and/or time." For particle counting measurements
(WBC, RBC, PLT) and related measurements (HB, HCT), dilutions of blood and recovery of expected
concentrations is the more appropriate description. For some portions of the CBC (i.e. MCV), simple
dilution is not effective.
It is the responsibility of analyzer manufacturers to define the Analytical Measuring Interval (AMI),
whereas it is the responsibility of the end user to verify such claims. AMI values are typically listed in
the manufacturer-supplied operator’s manual as a set of specifications and performance claims.
Parameter Linearity limits Visible Range Sample Tested range Error limits
WBC Commercial range 0.4 - 106.6 ± 0.3 or

0 - 100 100 - 150
(103/µL) Human blood 17.4 - 121.8 ± 5%
RBC Commercial range 0.23 - 8.1 ± 0.07 or

0-8 8 - 18
(106/µL) Human blood 0.98 - 9.83 ± 3%
HGB Commercial range 0.7 - 23.5 ± 0.3 or

0 - 26 26 - 30
(g/dL) Human blood 2.8 - 28 ± 3%
HCT Commercial range 2.0 - 71.2 ± 2 or

0 - 80 80 - 90
(%) Human blood 8.47 - 84.7 ± 3%
HGB > 2g/dL:

2200 - 6000 Human blood 221 - 2210
PLT 0 - 2200 ± 10 or
(103/µL) HGB < 2g/dL: ± 10%
4000 - 6000 Commercial range 9.75 - 4164
0 - 4000
Each obtained value must be within the range (defined by the Error limits) regarding the expected
value.
AMI validation is the process of confirming that the assay system will correctly recover the quantity of
the measurand over the AMI.
WBC Calibration Verification Accuracy & Linear Regression Plot

10
9
y=x
8 R2 = 1
7
Obtained result G/L
0
0 1 2 3 4 5 6 7 8 9
Expected result G/L
The more appropriate analytical term is recovery, as established by serial dilutions of a known high
concentration of cells. The goal is to establish the AMI, which may be a subset of the CRI, as follows:

Ref: RAL110BEN
■ The AMI is the range of analyte values that a method can directly measure on the specimen
without any dilution, concentration, or other pretreatment not part of the usual assay process.
■ The Clinically Reportable Interval (CRI) is the range of analyte values that a method can
measure, allowing for specimen dilution, concentration, or other pretreatment used to extend the
direct AMI. A manufacturer establishes the AMI through its validation testing whereas the CRI is
the end-user clinical laboratory’s responsibility.
White Blood Cell Concentration, Red Blood Cell Concentration, Hemoglobin, Hematocrit and
Platelet Concentration.
Commercially available "linearity kits" are assayed and evaluated. These data may be used to
represent the AMI based on commercial kit material processed per manufacturers’ directions, but do
not necessarily represent performance with actual fresh blood clinical specimens.
Clinically Reportable Intervals

The establishment of the CRI is a medical judgment made by the laboratory director to meet medical
needs, and is based in part on the assay technology. If the end-user laboratory verified the AMI and
the CRI is narrower, then no further action is required. Conversely, reporting patient results beyond
the AMI requires validation by the individual laboratory. The CRI is typically established at the time of
initial method verification and does not need re-evaluation unless the methodology changes.
For example, to report a result that exceeds the upper limit of the AMI, the specimen is commonly
diluted to bring the analyte into that range, the diluted specimen is reassayed, and the final result is
calculated using the dilution factor. The method manufacturer frequently specifies the AMI and
procedures to use for dilution or concentration of specimens with values outside the AMI. The lower
limit of the CRI is typically the lower limit of the AMI of the method, as verified during method
validation. Values lower than this limit may be reported as less than the limit. For hematology, it is
essential to understand that the lower limit of the AMI cannot extend lower than the LoB (Limit of
Blank); previous common claims of a "linearity" range to zero for PLT or WBC lack scientific sense if
the "background" is a nonzero value.
3.5.10. Contamination Between Samples
Contamination was assessed by running a sample with high results for the following parameters, three
times in a row (H1, H2, H3) followed by a sample with low results, also 3 consecutive times (L1, L2,
L3).
(L1 - L3)
Carry-over (%) = ________ x 100
(H3 - L3)
ABX Micros ES 60 OT
WBC (103/mm3) RBC (106/mm3) HGB (g/dL) PLT (103/mm3)

Mean concentration
0.98 0.98 3.30 38.17
of the lower level
Mean concentration
52.48 6.89 21.22 933.33
of the upper level
Contamination (%) 0.0 % 0.17 % 0.0 % 0.23%
Specifications <2% <2% <2% <2%
ABX Micros ES 60 CT

Mean concentration
0.90 0.98 3.12 32.17
of the lower level
Mean concentration
52.07 7.02 20.95 907.83
of the upper level

Ref: RAL110BEN

Contamination (%) 0.1 % 0.25 % 0.0 % 0.14 %
Specifications <2% <2% <2% <2%
Results obtained are within the specifications. There is no significant carry-over effect observed.
3.5.11. Stability
Reagent stability
Refer to the reagent leaflets for stability (Storage and Stability chapter). Latest version of documents
online at www.horiba.com.
3.5.12. Robustness
The robustness of an analysis method is the resistance to any variation in its results when minor
modifications occur regarding experimental conditions described in the procedure.
The inter-laboratory comparison (via Quality Control Program - QCP) gives a monthly overview of the
influence of factors such as: changing instrument, changing reagents, maintenance operations,
operator, geographic place, calibration, and temperature, humidity and pressure conditions.
Robustness of the blood count parameters can thereby be confirmed.
Results transmitted to QCP in October 2009 on control blood batches allowed the drawing up of the
following tables.
Precision of the peer-groups allows to detemine the robustness of the tests.
ABX Minotrol 16 on ABX Micros ES 60

Low level:
Number of CV inter- CV intra-instrument

Parameter
instruments instruments Min Mean Max
WBC 44 2.15 2.27 5.15 9.87
RBC 43 2.31 0.87 2.27 3.21
HGB 45 5.94 1.03 3.03 10.00
HCT 45 16.01 1.27 2.72 6.21
MCV 43 69.37 0.29 1.61 2.86
RDW 42 13.33 1.35 5.04 14.52
MCH 42 25.75 1.03 2.81 9.92
MCHC 42 37.10 0.98 2.93 9.81
PLT 45 89 4.48 9.31 19.42
MPV 42 8.71 1.16 4.69 11.22
LYM % 42 62.06 1.38 2.62 5.52
LYM # 39 1.29 1.98 6.39 15.19
MON % 42 10.72 4.51 9.08 29.61
MON # 39 0.20 10.00 12.85 49.36
GRA % 42 27.23 2.71 5.30 12.61
GRA # 39 0.69 5.71 10.34 32.90

Ref: RAL110BEN
Normal level:

Parameter
WBC 45 7.52 1.20 2.48 4.00
RBC 44 4.70 0.85 1.70 2.31
HGB 46 13.61 0.73 2.05 8.03
HCT 46 37.27 0.70 1.94 2.65
MCV 43 79.45 0.39 1.33 2.50
RDW 43 12.18 1.01 4.78 15.39
MCH 43 28.98 0.52 1.85 3.52
MCHC 43 36.50 0.57 1.99 4.23
PLT 46 272 2.12 5.26 7.94
MPV 43 8.01 0.86 3.24 12.66
LYM % 42 33.09 1.14 3.40 9.09
LYM # 41 2.44 2.00 5.06 12.00
MON % 43 6.65 3.75 8.71 35.71
MON # 41 0.44 5.92 16.32 50.00
GRA % 43 60.29 0.57 1.86 5.83
GRA # 41 4.63 1.28 3.07 7.78
High level:

Parameter
WBC 44 20.18 0.70 2.18 4.00
RBC 43 5.92 0.52 1.65 3.03
HGB 45 18.54 0.54 1.59 3.65
HCT 45 52.09 0.43 1.96 2.66
MCV 43 87.87 0.34 1.27 2.27
RDW 43 12.12 1.27 4.81 15.87
MCH 43 31.33 0.22 1.76 3.35
MCHC 43 35.60 0.39 1.97 4.27
PLT 45 493 1.35 4.58 5.45
MPV 43 7.86 1.00 2.79 12.66
LYM % 43 15.48 1.45 5.28 16.67
LYM # 41 3.07 1.65 6.33 18.33
MON % 42 4.38 0.43 19.61 95.00
MON # 39 0.82 2.50 20.28 75.00
GRA % 43 80.02 0.23 1.26 4.35
GRA # 41 16.25 0.62 2.38 4.66

Ref: RAL110BEN
3.5.13. Known Interfering Substances
3.5.13.1. Summary of Known Interferences
Parameter Causes of false increases Causes of false decreases

Unlysed red blood cells
Multiple myeloma
Hemolysis
Leukemia
WBC Platelet aggregation
Chemotherapy
Cryoglobulins
Macrothrombocytes
Erythroblasts
RBC agglutination
RBC Dilution of red blood cells
Cold agglutinins
Turbidity of the blood sample
HGB Increase in turbidity
Fetal blood
HCT RBC agglutination
MCV RBC agglutination
MCH Interferences cited for HGB and RBC
MCHC Interferences cited for HGB and HCT
Interferences cited for RBC and MCV
RDW Agglutination of red blood cells
Nutritional deficiency or blood transfusion
Very small erythrocytes
RBC agglutination
Hemolysis
Excessive numbers of giant platelets
Erythrocytic inclusions
PLT Chemotherapy
Elevated bilirubine
Platelet aggregation
Elevated lipids and/or cholesterol
A.C.D. blood
Parenteral nutrition
Giant platelets
MPV Very small erythrocytes RBC agglutination
Chemotherapy
LYM Presence of erythroblasts, certain parasites and erythrocytes
MON Presence of large lymphocytes, atypical lymphocytes, blasts, and excessive number of basophils
Presence of excessive numbers of eosinophils, metamyelocytes, myelocytes, promyelocytes,
NEU
blasts, and plasma cells
EOS Presence of abnormal granulations (degranulation of certain areas, toxic granulations, etc.)
Presence of significant numbers of WBC and / or LIC, as well as the contamination of the counting
BAS
channel
3.5.13.2. Interferences on White Blood Cells (WBC)
Unlysed red blood cells: in certain cases of membrane resistance, partial lysis of red blood cells may
be observed. These unlysed red blood cells may cause an erroneously high white blood cell count.
These unlysed red cells can be detected on the WBC curve via an L1 alarm or in the form of an
elevated base line of the lateral ascending section of the lymphocyte population.
Multiple myeloma: the precipitation of immunoglobulins in patients with multiple myeloma may give
elevated WBC counts.
Hemolysis: hemolyzed specimens contain an erythrocyte stroma, which may cause elevated white
blood cell counts.
Platelet agglutination: the accumulation of platelets may cause an elevated white blood cell count.
Platelet agglutination triggers the alarms L1.

Ref: RAL110BEN
Leukemia: leukemia can cause fragility of the leukocytes and subsequent destruction of these cells
during the count, thus resulting in an abnormally low white blood cell count. An abnormally low
leukocyte count may also be seen in patients with chronic lymphoblastic leukemia due to the
presence of abnormally small lymphocytes, which may not be counted by the analyzer.
Chemotherapy: cytotoxins and immunosuppressants may weaken the leukocyte membranes and
result in a low leukocyte count.
Cryoglobulins: the increased levels of cryoglobulins that may be associated with various conditions
(myeloma, carcinoma, leukemia, macroglobulinema, lymphoproliferative disorders, metastatic tumors,
autoimmune disorders, infections, aneurysms, pregnancy, thromboembolic phenomena, diabetes,
etc.), may cause an increase in the leukocyte, erythrocyte, and platelets counts and the hemoglobin
concentration. The samples should be warmed to 37°C (99°F) in a water bath for 30 minutes and then
re-run immediately afterwards (using the analyzer or a manual method).
Macrothrombocytes: in excessive numbers, they may affect the leukocyte count by increasing the
number of leukocytes counted.
Erythroblasts: high concentration of erythroblasts may increase the leukocyte count.
3.5.13.3. Interferences on Red Blood Cells (RBC)
The red blood cell dilution contains all of the elements found in the blood: erythrocytes, leukocytes,
and platelets. During the erythrocyte count, the platelets are not counted as they are smaller than the
defined minimum threshold. In very rare cases with an extremely high leukocyte count, the erythrocyte
count may be increased. It should be corrected, especially if the latter is very low in comparison with
the leukocyte count.
Agglutinated red blood cells: these may cause a falsely low RBC count. Blood samples containing
agglutinated red blood cells can be identified by abnormal MCH and MCHC values and the
examination of a stained blood smear.
Cold agglutinins: IgM, which are elevated in Cold Agglutinin Disease, may lower erythrocyte and
platelet counts and increase the MCV. The samples should be warmed to 37°C (99°F) in a water bath
for 30 minutes and then re-run immediately afterwards (using the analyzer or a manual method).
3.5.13.4. Interferences on Hemoglobin (HGB)
Turbidity of the blood sample: several physiological and/or therapeutic factors may produce falsely
elevated hemoglobin results. To obtain accurate results in blood samples with increased turbidity,
determine the cause of the turbidity and follow the appropriate method below:
■ An elevated leukocyte count: a very high leukocyte count will cause excessive diffusion of the
light. In such cases, the reference methods (manual) should be used. The diluted sample should
be centrifuged, and the supernatant fluid measured with a spectrophotometer.
■ Elevated lipemia: elevated lipemia levels makes the plasma look milky. This phenomenon can be
seen with hyperlipidemia, hyperproteinemia (as in gammopathies) and hyperbilirubinemia.
Accurate hemoglobin measurement can be achieved by using reference (manual) methods and a
plasma blank.
Increased turbidity: this phenomenon can be seen with red blood cells that are resistant to lysis. It
causes a falsely elevated HGB concentration, but can be detected due to abnormal MCHC and MCH
values and to an increase in the base line of the ascending section of the WBC curve. An erroneous
hemoglobin concentration also causes erroneous MCH and MCHC values.
Fetal blood: the mixing of fetal and maternal bloods may produce a falsely elevated hemoglobin
value.

Ref: RAL110BEN
3.5.13.5. Interferences on Hematocrit (HCT)
Red blood cells agglutination: can cause an inaccurate HCT value. Red blood cell agglutination may
be detected by observing abnormal MCV and MCH values, and by examining a stained blood smear.
In such cases, manual methods may be required to obtain an accurate hematocrit value.
3.5.13.6. Interferences on the Mean Cell Volume (MCV)
Red blood cell agglutination: can cause an inaccurate MCV value. Red blood cell agglutination may
be detected by observing abnormal MCH and MCHC values, and by examining a stained blood
smear.
Excessive numbers of large platelets: and/or the presence of an excessively high WBC count may
interfere with the accurate determination of the MCV value. In such cases, careful examination of a
stained blood smear may reveal the error.
3.5.13.7. Interferences on the Mean Corpuscular Hemoglobin (MCH)
The interferences cited for HGB and RBC affect the MCH and may cause inaccurate results.
3.5.13.8. Interferences on the Mean Corpuscular Hemoglobin Concentration

(MCHC)
The interferences cited for HGB and the HCT affect the MCHC and may cause inaccurate results.
3.5.13.9. Interferences on the Red Distribution Width (RDW)
The interferences cited for RBC and MCV affect the RDW and may cause inaccurate results.
Red blood cell agglutination: this phenomenon may cause a falsely low erythrocyte count and an
erroneous RDW. In the blood samples, red blood cell agglutination may be detected by observing
abnormal MCH and MCHC values, and by examining a stained blood smear.
Nutritional deficiency or blood transfusion: these phenomena may cause elevated RDW results due
to iron, vitamin B12, or folate deficiencies. It is also possible to observe an elevated RDW from the
bimodal distribution of red blood cells from transfused blood.
3.5.13.10. Interferences on Platelets (PLT)
Very small erythrocytes (microcytes): the presence of erythrocyte fragments (schistocytes), and
WBC fragments may interfere with the platelet count giving falsely elevated values.
Red blood cell agglutination: may trap the platelets and cause a falsely low platelet count. Red
blood cell agglutination may be detected by observing abnormal MCH and MCHC values, and by
examining a stained blood smear.
Excessive numbers of Macro platelets: this phenomenon may cause a falsely low platelet count
due to the fact that these macro platelets exceed the upper threshold defined for platelets and are
therefore not counted as platelets.
Chemotherapy: cytotoxins and immunosuppressants may weaken these cells and result in a falsely
low count. Manual methods may be necessary to obtain the platelet count.
Hemolysis: hemolyzed samples contain a red blood cell stroma which may affect the platelet count.

Ref: RAL110BEN
Citrate blood - Blood anti-coagulated with citrate may contain platelet aggregates which could
decrease the platelet count.
RBC Inclusions: including Howell-Jolly bodies, Heinz bodies, siderotic and basophilic granules, etc.,
may cause falsely elevated platelet counts.
Platelet agglutination: the accumulation of platelets may cause a low platelet count. The sample
should be repeated and drawn into a sodium citrate anticoagulant tube to rule out the anticoagulant
as a cause of aggregation and run again to determine the platelet count alone. The final platelet count
should be corrected, making allowance for the dilution caused by the sodium citrate. Platelet
agglutination triggers the alarms L1.
Elevated lipids and/or cholesterol: may interfere with correct platelet counting. From patients
undergoing parenteral nutrition with intralipids brought, it is noted an over-estimate of the platelet
counting which can mask a thrombopenia.
Elevated bilirubine: may interfere with correct platelet counting. From patients with severe hepatic
disorder, liver transplant… It is noted an over-estimate of the platelet counting which can mask a
thrombopenia.
Parenteral nutrition: Interference in PLT result may occur for samples from patients undergoing
parenteral nutrition with injection of lipid emulsion.
3.5.13.11. Interferences on the Mean Platelet Volume (MPV)
Macro platelets: their volume exceeds the upper threshold defined for platelets and they are not
therefore included in the calculation of the mean platelet volume by the analyzer. The MPV value may
be falsely lowered.
Very small erythrocytes (microcytes) or presence of red blood cell fragments (schistocytes) and
white blood cell fragments may interfere with the accurate determination of the mean platelet
volume.
Red blood cell agglutination: may trap the platelets causing an incorrect MPV. Red blood cell
agglutination may be detected by observing abnormal MCH and MCHC values, and by examining a
stained blood smear.
Chemotherapy: may also affect platelet volume.
Blood samples collected in EDTA will not maintain a stable Mean Platelet Volume.
Platelets collected in EDTA swell with time and temperature.
3.5.13.12. Interferences on Lymphocytes (LYM)
The presence of erythroblasts and erythrocytes that are resistant to lysis may cause an inaccurate
lymphocyte count. Limitations to the leukocyte count also apply to the determination of the number
(absolute value) and percentage of lymphocytes.
3.5.13.13. Interferences on Monocytes (MON)
The presence of large lymphocytes, atypical lymphocytes, lymphoblasts, and excessive numbers of
basophils may cause an inaccurate monocyte count. Limitations to the leukocyte count also apply to
the determination of the number (absolute value) and percentage of monocytes.

Ref: RAL110BEN
3.5.13.14. Interferences on Granulocytes (GRA)
The excessive presence of eosinophils, metamyelocytes, promyelocytes, myeloblasts, myelocytes

and plasma cells may cause an inaccurate determination of the number (absolute value) and
percentage of granulocytes (GRA).
3.5.13.15. Interfering Substances
This study is based on known interferences on the predicate device (ABX Micros 60) and on the CLSI
EP7-A2 – Interference testing in clinical chemistry.
Interferent test concentration
The paired interference testing is conducted at the highest concentrations that a laboratory would
expect to observe among patient specimen.
Potential Interfering substances:
■ Hemolysis
■ Icterus (total bilirubin)
■ Urea
■ Lipemia
Acceptance Criteria
Evaluating the effect of potentially interfering substances added to the sample of interest
The smallest significant differences between test and control is:
%
WBC RBC HGB HCT MCV MCH MCHC RDW PLT MPV % LYM % GRA
MON
Relative
7% 5% 4% 6% 2% 4% 4.3% 9% 22% 9% 1.3% 30% 7%
%
_ _
Compute the observed interference effect, dobs = Interference = (X test - X control)
A one sided test is performed. The 95 % confidence interval for the interference effect is calculated. If
the point estimate dobs is less than or equal to the cut-off value, dc, conclude the bias caused by the
substance is less than dmax.
Urea
On all parameters, no significant interference can be observed with urea for a concentration of
29.5 mmol/L, either on low, normal or high level.
Bilirubin
On all parameters, no significant interference can be observed with bilirubin for a concentration of
150 µmol/L, either on low, normal or high level.
Intra Lipid
On all parameters, no significant interference can be observed with Intra Lipid for a concentration of
3 mmol/L, either on low, normal or high level.
Hemolysis
On all parameters, no significant interference can be observed with Hemolysis. The WBC value is
slightly increased but still into the acceptance criteria.

Ref: RAL110BEN
3.5.14. Reference Values
The reference values vary according to the population and / or the region. Each laboratory is strongly
advised to establish its own set of normal ranges according to the local population.
Parameters Male Female

WBC (103/mm3) 4.00 - 10.00 4.00 - 10.00
RBC (106/mm3) 4.50 - 6.50 3.80 - 5.80
HGB (g/dL) 13.00 - 17.00 11.50 - 16.00
HCT (%) 40 - 54 37 - 47
MCV (fL) 80.00 - 100.00 80.00 - 100.00
MCH (pg) 27.00 - 32.00 27.00 - 32.00
MCHC (g/dL) 32.00 - 36.00 32.00 - 36.00
RDW (%) 11 - 16 11 - 16
PLT (103/mm3) 150 - 500 150 - 500
MPV (µm3) 6.00 - 11.00 6.00 - 11.00
LYM (103/mm3) 1.00 - 5.00 1.00 - 5.00
MON (103/mm3) 0.20 - 1.50 0.20 - 1.50
NEU (103/mm3) 2.00 - 8.00 2.00 - 8.00
EOS (103/mm3) 0.00 - 0.70 0.00 - 0.70
BAS (103/mm3) 0.00 - 0.25 0.00 - 0.25
WBC (103/mm3):
Male Female
Age
# of individuals Reference values # of individuals Reference values
4 - 10 1 880 4.30 - 11.90 1 755 4.40 - 11.70
10 - 18 1 800 4.10 - 10.80 1 620 4.20 - 11.60
18 - 65 3 317 4.10 - 10.50 2 709 6.30 - 10.50
RBC (106/mm3):
Male Female
Age
4 - 10 1 877 4.20 - 5.30 1 752 4.10 - 5.30
10 - 14 1 167 4.30 - 5.50 168 4.20 - 5.30
14 - 18 1 627 4.50 - 5.70 1 548 4.20 - 5.20
18 - 25 1 562 4.60 - 5.80 1 576 4.10 - 5.20
25 - 45 2 209 4.50 - 5.70 232 4.10 - 5.20
45 - 65 1 509 4.40 - 5.60 186 4.10 - 5.40
HGB (g/dL):
Male Female
Age
4 - 10 1 878 11.6 - 14.8 1 754 11.6 - 14.9
10 - 14 1 170 12.5 - 15.8 170 12.4 - 15.3
14 - 18 1 631 13.5 - 17.4 1 550 12.6 - 15.6
18 - 25 1 571 14.1 - 17.8 1 574 12.4 - 15.7
25 - 35 1 480 14.1 - 17.6 1 175 11.9 - 15.6

Ref: RAL110BEN
Male Female
Age
35 - 45 1 744 14.0 - 17.5 1 566 11.6 - 15.8
45 - 55 1 341 13.7 - 17.4 1 291 11.5 - 15.8
55 - 65 1 173 13.6 - 17.6 186 12.7 - 15.9
MCV (fL):
Male Female
Age
4 - 10 1 858 76 - 88 1 740 76 - 89
10 - 14 1 158 78 - 90 148 79 - 91
14 - 18 1 624 80 - 95 1 544 80 - 95
18 - 25 1 571 81 - 97 1 571 81 - 96
25 - 35 1 487 83 - 97 1 178 80 - 97
35 - 45 1 744 83 - 98 1 569 77 - 98
45 - 55 1 340 84 - 98 1 294 76 - 97
55 - 65 1 172 84 - 100 186 81 - 99
PLT (103/mm3):
Male Female
Age
4 - 10 7 215 203 - 484 6 726 210 - 482
10 - 14 5 211 185 - 419 4 882 186 - 422
14 - 18 4 163 165 - 385 4 068 175 - 390
18 - 25 2 730 150 - 349 3 934 164 - 384
25 - 45 12 273 153 - 355 13 601 161 - 386
45 - 55 3 515 152 - 368 3 760 168 - 399
55 - 65 1 367 151 - 365 1 480 171 - 386
> 65 260 143 - 361 306 154 - 386
Polynuclear neutrophils (103/mm3):
Male Female
Age
4 - 10 1 880 1.30 - 8.70 1 755 1.40 - 9.00
10 - 18 1 800 1.40 - 8.10 1 620 1.60 - 8.40
18 - 65 3 317 1.70 - 8.00 2 709 1.70 - 8.20
Polynuclear eosinophils (103/mm3):
Male Female
Age
4 - 10 1 880 0 - 1.20 1 755 0 - 1.10
10 - 18 1 800 0 - 1.90 1 620 0 - 1.90
18 - 65 3 317 0 - 1.70 2 709 0 - 1.70

Ref: RAL110BEN
Polynuclear basophils (103/mm3):
Male Female
Age
4 - 10 1 880 0 - 0.40 1 755 0 - 0.40
10 - 18 1 800 0 - 0.30 1 620 0 - 0.30
18 - 65 3 317 0 - 0.30 2 709 0 - 0.30
Lymphocytes (103/mm3):
Male Female
Age
4 - 10 1 880 1.90 - 7.10 1 755 1.00 - 7.10
10 - 18 1 800 1.80 - 6.10 1 620 1.80 - 6.10
18 - 65 3 317 1.70 - 5.20 2 709 1.70 - 5.30
Monocytes (103/mm3):
Male Female
Age
4 - 10 1 880 0.02 - 1.30 1 755 0.02 - 1.20
10 - 18 1 800 0.03 - 1.30 1 620 0.02 - 1.20
18 - 65 3 317 0.04 - 1.30 2 709 0.02 - 1.10
References:
AIDE MEMOIRE D’HEMATOLOGIE, 1998. Prof. C. Sultan / M. Gouault - Helman / M. Imbert, Service
Central d’Hématologie de l’Hôpital Henri Mondor, Faculté de médecine de Créteil (Paris XII).
Lecture critique de l’hémogramme : valeurs seuils à reconnaître comme probablement pathologiques
et principales variations non pathologiques. ANAES : Agence Nationale d’Accréditation et d’Evaluation
en Santé.
3.5.15. Comparison with a Reference Instrument
The correlation of the following parameters of the analyzer has been proven by regression analysis
comparing the analyzer with two recognized comparative analyzers, on 100 samples of whole blood
from patients, within the normal operating ranges of the analyzer (according to EP09-A2 evaluation
protocol):
ABX Micros ES 60 OT
R2 (comparison of means)
Parameter Accuracy requirements
Versus ABX Micros 60 Versus ABX Pentra 80
WBC 0.9971 0.9947 > 0.95
RBC 0.9915 0.9970 > 0.95
HGB 0.9951 0.9981 > 0.95
HCT 0.9909 0.9942 > 0.95
MCV 0.9884 0.9909 > 0.95
PLT 0.9826 0.9868 > 0.95
LYM% 0.9881 - > 0.95
MON% 0.9205 - > 0.95

Ref: RAL110BEN
GRA% 0.9898 - > 0.95
ABX Micros ES 60 CT
WBC 0.9963 0.9952 > 0.95
RBC 0.9873 0.9928 > 0.95
HGB 0.9879 0.9919 > 0.95
HCT 0.9858 0.9849 > 0.95
MCV 0.9647 0.9662 > 0.95
PLT 0.9860 0.9863 > 0.95
LYM% 0.9890 - > 0.95
MON% 0.8680 - > 0.95
GRA% 0.9885 - > 0.95
ABX Micros 60 ABX Micros ES 60 OT ABX Micros ES 60 CT ABX Micros 60

Min 1.50 1.55 Min 1.49 1.50
Max 15.85 16.20 Max 16.38 15.85
Mean 7.11 7.10 Mean 7.06 7.11

Ref: RAL110BEN
ABX Pentra 80 ABX Micros ES 60 OT ABX Micros ES 60 CT ABX Pentra 80

Min 1.62 1.55 Min 1.50 1.62
Max 16.11 16.20 Max 16.55 16.11
Mean 7.11 7.10 Mean 7.13 7.11
ABX Micros 60 ABX Micros ES 60 OT ABX Micros ES 60 ABX Micros 60

CT
Min 2.30 2.32 Min 2.23 2.27
Max 5.80 5.77 Max 5.70 5.71
Mean 4.05 4.05 Mean 3.99 3.99

Ref: RAL110BEN
ABX Pentra 80 ABX Micros ES 60 OT ABX Micros ES 60 CT ABX Pentra 80

Min 2.28 2.32 Min 2.23 2.28
Max 5.72 5.77 Max 5.70 5.72
Mean 4.01 4.05 Mean 3.99 4.01
ABX Micros 60 ABX Micros ES 60 OT ABX Micros ES 60 CT ABX Micros 60

Min 7.02 7.15 Min 6.75 6.89
Max 16.69 16.75 Max 16.10 16.37
Mean 12.38 12.40 Mean 12.11 12.16

Ref: RAL110BEN
ABX Pentra 80 ABX Micros ES 60 OT ABX Micros ES 60 ABX Pentra 80

CT
Min 6.82 7.15 Min 6.75 6.62
Max 17.11 16.75 Max 16.10 16.61
Mean 12.45 12.42 Mean 12.11 12.08

CT
Min 112.79 120 Min 110 109.5
Max 640.15 674.5 Max 647 621.5
Mean 265.84 268.69 Mean 256.49 258.1

Ref: RAL110BEN

CT
Min 112.20 120 Min 110 110
Max 677.28 674.5 Max 647 664
Mean 265.39 268.69 Mean 256.49 260.08

CT
Min 20.29 20.45 Min 19.65 19.90
Max 50.16 49.55 Max 48.40 49.19
Mean 37.10 37.03 Mean 36.38 36.38

Ref: RAL110BEN

CT
Min 20.68 20.45 Min 19.65 20.28
Max 49.83 49.55 Max 48.40 48.85
Mean 37.07 37.03 Mean 36.38 36.34
3.6. Labeling

Each item of equipment shall be uniquely labelled, marked or otherwise identified.
Refer to the Introduction > Labels and Connections chapter of the user manual (RAB237) of the
ABX Micros ES 60.

Ref: RAL110BEN
3.7. Individual Machine Data Sheets

Records shall be maintained for each item of equipment contributing to the performance of
examinations. These records shall include at least the following:
■ a- identity of the equipment;
■ b- manufacturer's name, type identification and serial number or other unique identification;
■ c- manufacturer's contact person and telephone number, as appropriate;
■ d- date of receiving and date of putting into service;
■ e- current location, where appropriate;
■ f- condition when received (e.g. new, used or reconditioned);
■ g- manufacturer's instructions, if available, or reference to their retention;
■ h- equipment performance records that confirm the equipment's suitability for use;
■ i- maintenance carried out and that planned for the future;
■ j- damage to, or malfunction, modification or repair, of the equipment;
■ k- predicted replacement date, if possible.
The performance records referred to in h) should include copies of reports/certificates of all
calibrations and/or verifications including dates, time and results, adjustments, the acceptance criteria
and due date of the next calibration and/or verification, together with the frequency of checks carried
out between maintenance/calibration, as appropriate, to fulfil part or all of this requirement.
Manufacturer's instructions may be used to establish acceptance criteria, procedures and frequency of
verification for maintenance or calibration or both, as appropriate, to fulfil part or all of this
requirement. These records shall be maintained and shall be readily available for the life span of the
equipment or for any time period required by national, regional and local regulations.
Refer to the Quality Management System chapter of this document.
3.8. Manuals and Instructions for Use

Equipment shall be operated by authorized personnel only. Up-to-date instructions on the use and
maintenance of equipment (including any relevant manuals and directions for use provided by the
manufacturer of the equipment) shall be readily available to laboratory personnel.
HORIBA Medical provides an electronic version or a paper version (on demand) of the user manual for
ABX Micros ES 60:
The reference of the current version of the ABX Micros ES 60 software is given in the revision table of
the Foreword section. HORIBA Medical provides reagent leaflets and Material Safety Data Sheets for
reagents, controls and calibrators in a multilingual electronic version. These documents are available
on the CD-Rom RAX055X (X being the revision index of the CD-Rom) which is supplied with the
instrument. These documents are also available online at www.horiba.com.

Ref: RAL110BEN
3.9. Disposal of Chemical Products and Biological Materials

Equipment shall be maintained in a safe working condition. This shall include examination of
electrical safety, emergency stop devices and the safe handling and disposal of chemical,
radioactive and biological materials by authorized persons. Manufacturers' specifications or
instructions or both shall be used, as appropriate.
Refer to the Waste Disposal chapter of this document.
3.10. Use of Computers for the Analysis Data

When computers or automated examination equipment are used for the collection, processing,
recording, reporting, storage or retrieval of examination data, the laboratory shall ensure that:
■ a- computer software, including that built into equipment, is documented and suitably validated as
adequate for use in the facility;
■ b- procedures are established and implemented for protecting the integrity of data at all times;
■ c- computers and automated equipment are maintained to ensure proper functioning and provided
with environmental and operating conditions necessary for maintaining the integrity of data;
■ d- computer programmes and routines are adequately protected to prevent access, alteration or
destruction by casual or unauthorized persons.
Use and protection of computer programmes

ABX Micros ES 60 has an integrated computer allowing to get and save results.
The access to this computer is protected by a login and a password.
Validation and software documentation

The ABX Micros ES 60 includes a software programme. This instrument is a in vitro diagnostic
medical device with the CE mark and cleared by the US FDA. In accordance with the applicable
regulation for this product, its integrated software programme has been designed to ensure its
repeatability, reliability and performance, in accordance with its intended use. Horiba Medical has
developed and implemented a verification and validation process for its instruments.
Based in particular on the requirements of the 21 CFR Part 11: Electronic Records; Electronic
Signatures (North American regulations), integrated software programmes are protected to prevent
access, alteration or deletion of data by casual or unauthorized persons.

Ref: RAL110BEN
Pre-analytical Procedures
4. Pre-analytical Procedures

The primary sample collection manual shall include the following:
■ a- copies of or references to:
■ 1- lists of available laboratory examinations offered,
■ 2- consent forms, when applicable,
■ 3- information and instructions provided to patients in relation to their own preparation
before primary sample collection,
■ 4- information for users of laboratory services on medical indications and appropriate
selection of available procedures;
■ b- procedures for:
■ 1- preparation of the patient (e.g. instructions to caregivers and phlebotomists),
■ 2- identification of primary sample,
■ 3- primary sample collection (e.g. phlebotomy, skin puncture, blood, urine and other body
fluids), with descriptions of the primary sample containers and any necessary additives;
■ c- instructions for:
■ 1- completion of request form or electronic request,
■ 2- type and amount of the primary sample to be collected,
■ 3- special timing of collection, if required,
■ 4- any special handling needs between time of collection and time received by the
laboratory (transport requirements, refrigeration, warming, immediate delivery, etc.),
■ 5- labelling of primary samples,
■ 6- clinical information (e.g. history of administration of drugs),
■ 7- positive identification, in detail, of the patient from whom a primary sample is collected,
■ 8- recording the identity of the person collecting the primary sample,
■ 9- safe disposal of materials used in the collection;
■ d- instructions for:
■ 1- storage of examined samples,
■ 2- time limits for requesting additional examinations,
■ 3- additional examinations,
■ 4- repeat examination due to analytical failure or further examinations of same primary
sample.
4.1. Specimen for Sampling
Refer to the Specifications > Limitations chapter of the user manual (RAB237) of the
ABX Micros ES 60.

Ref: RAL110BEN
Analytical Procedures
5. Analytical Procedures

All procedures shall be documented and be available at the workstation for relevant staff.
Documented procedures and necessary instructions shall be available in a language commonly
understood by the staff in the laboratory.
Card files or similar systems that summarize key information are acceptable for use as a quick
reference at the workbench, provided that a complete manual is available for reference. The card
file or similar systems shall correspond to the complete manual. Any such abridged procedures
shall be part of the document control system.
The procedure shall be based on the instructions for use (e.g. package insert) written by the
manufacturer, provided that they are in accordance with 5.5.1 and 5.5.2 and that they describe the
procedure, as it is performed in the laboratory, and are written in the language commonly
understood by the staff of the laboratory. Any deviation shall be reviewed and documented.
Additional information that could be required to perform the examination shall also be
documented. Each new version of examination kits with major changes in reagents or procedure
shall be checked for performance and suitability for intended use. Any procedural changes shall be
dated and authorized as for other procedures.

Ref: RAL110BEN
Ensuring Quality of Examination Procedures
6. Ensuring Quality of Examination

Procedures

The laboratory shall design internal quality control systems that verify the attainment of the
intended quality of results. It is important that the control system provide staff members with clear
and easily understood information on which to base technical and medical decisions. Special
attention should be paid to the elimination of mistakes in the process of handling samples,
requests, examinations, reports, etc.
The laboratory shall determine the uncertainty of results, where relevant and possible. Uncertainty
components which are of importance shall be taken into account. Sources that contribute to
uncertainty may include sampling, sample preparation, sample portion selection, calibrators,
reference materials, input quantities, equipment used, environmental conditions, condition of the
sample and changes of operator.
A programme for calibration of measuring systems and verification of trueness shall be designed
and performed so as to ensure that results are traceable to SI units or by reference to a natural
constant or other stated reference. Where none of these is possible or relevant, other means for
providing confidence in the results shall be applied, including but not limited to the following:
■ a- participation in a suitable programme of interlaboratory comparisons;
■ b- use of suitable reference materials, certified to indicate the characterization of the material;
■ c- examination or calibration by another procedure;
■ d- ratio or reciprocity-type measurements;
■ e- mutual consent standards or methods which are clearly established, specified, characterized
and mutually agreed upon by all parties concerned;
■ f- documentation of statements regarding reagents, procedures or the examination system
when traceability is provided by the supplier or manufacturer.
Calibration
The controls and calibrators associated with the ABX Micros ES 60, supplied by HORIBA Medical,
comply with the requirements of the standard ISO 17511 : 2003, relating to the metrological
traceability of values attributed to calibration agents and control equipment (in vitro diagnostic
medical devices) for the measurement of quantities in biological samples. This makes it possible to
guarantee that the results obtained by the ABX Micros ES 60, duly controlled and calibrated, are
sufficiently exact to enable a clinically correct interpretation and comparability over time and space.
Traceability & Uncertainty

ABX Minocal Hematology Calibrator
Instrument Parameter Uncertainty Unit

WBC ± 0.2 103/µL
RBC ± 0.06 106/µL
HGB ± 0.2 g/dL
ABX Micros ES 60 HCT ± 1.0 %
MCV ± 2.0 fL
PLT ± 10 103/µL
MPV ± 0.5 fL

Ref: RAL110BEN
HORIBA Medical Hematology Control and Calibrator values, manufactured by R&D Systems, Inc are
traceable to standard reference methods.
Hematology analyzers in R&D Systems’ Quality Assurance Laboratory are whole blood calibrated to
values obtained using the following standard reference methods. Whole blood samples drawn from
normal, healthy donors are collected in EDTA anticoagulant and analyzed within six hours of
collection.
The White Blood Cell (WBC) and Red Blood Cell (RBC) are analyzed on a Coulter Counter Z series
instrument. All counts are corrected for coincidence.
Hemoglobin is measured using the Clinical Laboratory Standards Institute (CLSI) recommended
reagent for the hemoglobincyanide (cyanmethemoglobin) method. Readings are made at 540 nm in a
colorimeter/spectrophotometer calibrated according to CLSI H15-A3 and ICSH recommendations 7 .
The hematocrit (packed cell volume) is measured using plain glass microhematocrit tubes (not
coated with anticoagulant) centrifuged for 5 minutes in a microhematocrit centrifuge according to the
CLSI H7-A3 document 8 . No correction is made for trapped plasma.
Platelets are assayed using a hemocytometer and phase contrast optics.
All brands and products are trademarks or registered trademarks of their respective owners.
7 National Committee for Clinical Laboratory Standards (now Clinical Laboratory Standards Institute.) Reference and Selected
Procedures for the Quantitative Determination of Hemoglobin in Blood: Approved Standard-Third Edition. NCCLS document H15-
A3. Wayne, PA: NCCLS, 2000.
8 National Committee for Clinical Laboratory Standards (now Clinical Laboratory Standards Institute.) Procedure for Determining
Packed Cell Volume by the Microhematocrit Method: Approved Standard, NCCLS document H7-A3. NCCLS, Wayne, PA: NCCLS,
2001.

Ref: RAL110BEN
6.1. Traceability Chain for WBC and RBC Count
Primary reference
measurement
procedure
ICSH reference
method for the
enumeration of
RBC and WBC*
Manufacturer's
working
calibrator
Fresh human blood
Manufacturer's
standing measurement
procedure
ABX Minocal assay

determination
procedure on
standard instrument
Manufacturer's
product
calibrator
ABX Minocal
End user's
routine measurement
procedure
ABX Micros ES60
Routine sample
EDTA whole blood
Result
RBC and WBC count
* The White Blood Cell (WBC) and Red Blood Cell (RBC) are analyzed on a Coulter Counter Z series
instrument. All counts are corrected for coincidence.

Ref: RAL110BEN
6.2. Traceability Chain for HGB Measurement
Primary reference
measurement
procedure
ICSH reference
method for the
measurement of
Hemoglobin*
Manufacturer's
working
calibrator
Fresh human blood
Manufacturer's
procedure
ABX Minocal assay

determination
procedure on
standard instrument
Manufacturer's
product
calibrator
ABX Minocal
End user's
routine measurement
procedure
ABX Micros ES60
Routine sample
EDTA whole blood
Result
Hemoglobin
* Hemoglobin is measured using the Clinical Laboratory Standards Institute (CLSI) recommended
reagent for the hemoglobincyanide (cyanmethemoglobin) method(1). Readings are made at 540 nm in
a colorimeter/spectrophotometer calibrated according to CLSI H15-A3 and ICSH recommendations 9 .
9 National Committee for Clinical Laboratory Standards (now Clinical Laboratory Standards Institute.) Reference and Selected
Procedures for the Quantitative Determination of Hemoglobin in Blood: Approved Standard-Third Edition. NCCLS document H15-
A3. Wayne, PA: NCCLS, 2000.

Ref: RAL110BEN
6.3. Traceability Chain for HCT Measurement
Primary reference
measurement
procedure
ICSH reference
method for the
measurement of
Hematocrit*
Manufacturer's
working
calibrator
Fresh human blood
Manufacturer's
procedure
ABX Minocal assay

determination
procedure on
standard instrument
Manufacturer's
product
calibrator
ABX Minocal
End user's
routine measurement
procedure
ABX Micros ES60
Routine sample
EDTA whole blood
Result
Hematocrit
* The hematocrit (packed cell volume) is measured using plain glass microhematocrit tubes (not
coated with anticoagulant) centrifuged for five minutes in a microhematocrit centrifuge according to
the CLSI H7-A3 document 10 . No correction is made for trapped plasma.
10 National Committee for Clinical Laboratory Standards (now Clinical Laboratory Standards Institute.) Procedure for Determining
Packed Cell Volume by the Microhematocrit Method: Approved Standard, NCCLS document H7-A3. NCCLS, Wayne, PA: NCCLS,
2001.

Ref: RAL110BEN
6.4. Traceability Chain for PLT Count
Primary reference
measurement
procedure
ICSH reference
method for the
enumeration of
Platelets*
Manufacturer's
working
calibrator
Fresh human blood
Manufacturer's
procedure
ABX Minocal assay

determination
procedure on
standard instrument
Manufacturer's
product
calibrator
ABX Minocal
End user's
routine measurement
procedure
ABX Micros ES60
Routine sample
EDTA whole blood
Result
PLT count
* Platelets are assayed using a hemocytometer and phase contrast optics.

Ref: RAL110BEN
Results Report
7. Results Report
■ § 5.8 of the standard ISO 15189 : 2007

Refer to the Workflow section of the user manual (RAB237) of the ABX Micros ES 60.
Refer to the Settings section of the user manual (RAB237) of the ABX Micros ES 60.

Ref: RAL110BEN
Results Report

Ref: RAL110BEN
Appendices
1. Glossary of Terms.................................................................................................................68
2. References...............................................................................................................................78
2.1. Regulatory References..............................................................................................................78
2.2. General Standardization References........................................................................................78
2.3. COFRAC-EA Documentation....................................................................................................79
2.4. Method Validation.....................................................................................................................79
2.5. Websites...................................................................................................................................80

Ref: RAL110BEN
Appendices
Glossary of Terms
1. Glossary of Terms
Acceptability, Criteria of (Performance Standard)

Criteria according to which the performance of a technique are deemed satisfactory under the
conditions of use defined by the user (these criteria are primarily based on the concepts of
imprecision, inexactitude, and total allowable error).
Accuracy
Ability of the instrument to agree with a predetermined reference value at any point within the
operating range; closeness of a result to the true (accepted) value.
Analysis (Field of)

Interval of concentrations (or other quantities) of an analyte for which the technique is applicable
without modification. Its evaluation requires the establishment of linearity limits and (possibly) of the
detection limit of the technique. Synonym: “Field of measurement, range of measurement”.
Analyte
Component, substance, material to be measured in a possibly complex environment.
Analytical sensitivity
In compliance with the Common Technical Specifications (CTS), the «analytical sensitivity» refers to
the limit of detection, i.e. the smallest quantity of target marker that can be detected with precision.
Analytical specificity
The capacity of the method to determine only the target marker.
Background count
Measure of the amount of electrical or particle interference.
Bias (ISO 3534-1)

Difference between the mathematical prediction of the results of the analysis and the accepted
reference value.
Calibration
Set of operations to establish, under specified conditions, the relationship between the values of the
quantity indicated by a measuring instrument or a measurement system or the values represented by
a materialized measurement or by a reference material, and the corresponding values of the quantity
given by standards.
Calibration factors
These are correction factors that the system uses to fine-tune instrument accuracy.

Ref: RAL110BEN
Appendices
Glossary of Terms
Calibrator
A (reference) material (e.g., solution, suspension) or device of known quantitative/qualitative
characteristics (e.g., concentration, activity, intensity, reactivity) used to calibrate, graduate, or adjust
a measurement procedure or to compare the response obtained with the response of a test
specimen/sample (NCCLS H38-P)
Carry-over
Amount of blood cells remaining in diluent following the cycling of a blood sample (in percent).
Cell control
Preparation made of human blood with stabilized cells and surrogate material used for daily
instrument quality control.
Certified reference material

Reference material, accompanied by a certificate, of which one (or several) value(s) of the property(ies)
is (are) certified by a procedure that establishes its association with an exact undertaking of the unit in
which the property values are expressed and for which each certified value is accompanied by an
uncertainty with a known level of confidence.
Characterization (evaluation)
Study To test the analysis protocol (analytical procedure) to determine the values of performance
criteria, which are determined beforehand. This study can be undertaken on an intra-laboratory (by
the supplier or manufacturer) or inter-laboratory basis.
Chemical specificity, specificity

Property of an analytical method to selectively determine the concentration of the component(s) that it
is designed to measure.
Coefficient of variation (CV) ISO 3534-1

For a non-negative character, ratio of the standard deviation to the mean.
COFRAC
«Comité français d'accréditation» (French Accreditation Committee)
Mission: To attest that the accredited bodies are competent and impartial, to obtain on an
international level the acceptance of their services and the recognition of expertise by laboratories,
and inspection and certification bodies.
Accreditation bodies affiliated to the multilateral agreement, ILAC (International Laboratory
Accreditation Cooperation):
Organismo Argentino de Acreditacion (OAA), Argentina

National Association of Testing Authorities, Australia (NATA), Australia
Bundesministerium fur Wirtschaft und Arbeit (BMWA), Austria
BELAC (Belgian Accreditation Structure), Belgium
Coordenação Geral de Credenciamento General Coordination for Accreditation (CGCRE/INMETRO),
Brazil
Canadian Association for Environmental Analytical Laboratories (CAEAL), Canada
Standards Council of Canada (SCC), Canada
Hong Kong Accreditation Service (HKAS), Hong Kong, China
China National Accreditation Service for Conformity Assessment (CNAS), People’s Republic of China
Ente Costarricense de Acreditation (ECA), Costa Rica
National Accreditation Body of Republica de Cuba (ONARC), Cuba
Czech Accreditation Institute (CAI), Czech Republic
Danish Accreditation (DANAK), Denmark

Ref: RAL110BEN
Appendices
Glossary of Terms
National Laboratories Accreditation Bureau (NLAB), Egypt

Finnish Accreditation Service (FINAS), Finland
Comite Francais d’Accreditation (COFRAC), France
Deutsche Akkreditierungsstelle DACH, Germany
Deutsches Akkreditierungssystem Prüfwesen (DAP), Germany
Deutsche Akkreditierungsstelle Technik in Trägergemeinschaft für Akkreditierung German Association
for Accreditation GmbH (DATech in TGA GmbH), Germany
Deutscher Kalibrierdienst (DKD), Germany
Hellenic Accreditation System S.A. (ESYD), Greece
National Accreditation Board for Testing & Calibration Laboratories (NABL), India
National Accreditation Body of Indonesia (KAN), Indonesia
Irish National Accreditation Board (INAB), Ireland
Israel Laboratory Accreditation Authority (ISRAC), Israel
Sistema Nazionale per l’Accreditamento di Laboratori (SINAL), Italy
Servizio di Taratura in Italia (SIT), Italy
International Accreditation Japan (IA Japan), Japan
Japan Accreditation Board for Conformity Assessment (JAB), Japan
Voluntary EMC Laboratory Accreditation Center INC (VLAC), Japan
Korea Laboratory Accreditation Scheme (KOLAS), Republic of Korea
Department of Standards Malaysia (STANDARDS MALAYSIA), Malaysia
entidad mexicana de acreditación, a.c. (ema), Mexico
Dutch Accreditation Council (RvA), Netherlands
International Accreditation New Zealand (IANZ), New Zealand
Norsk Akkreditering (NA), Norway
Philippine Accreditation Office (PAO), Philippines
Polish Centre for Accreditation (PCA), Poland
Instituto Portugues de Acreditacao (IPAC), Portugal
Romanian Accreditation Association (RENAR), Romania
Singapore Accreditation Council (SAC), Singapore
Slovak National Accreditation Service (SNAS), Slovakia
Slovenian Accreditation (SA), Slovenia
South African National Accreditation System (SANAS), South Africa
Entidad Nacional de Acreditacion (ENAC), Spain
Swedish Board for Accreditation and Conformity Assessment (SWEDAC), Sweden
Swiss Accreditation Service (SAS), Switzerland
Taiwan Accreditation Foundation (TAF), Chinese Taipei
The Bureau of Laboratory Quality Standards, Department of Medical Sciences, Ministry of Public
Health, Thailand (BLQS-DMSc), Thailand
Thai Industrial Standards Institute (TISI), Thailand
Bureau of Laboratory Accreditation, Department of Science Service, Ministry of Science and
Technology (BLA-DSS), Thailand
Turkish Accreditation Agency (TURKAK), Turkey
United Kingdom Accreditation Service (UKAS), United Kingdom
American Association for Lab Accreditation (A2LA), USA
Assured Calibration and Laboratory Accreditation Select Services (ACLASS), USA
International Accreditation Service, Inc (IAS), USA
National Voluntary Laboratory Accreditation Program (NVLAP), USA
Laboratory Accreditation Bureau (L-A-B), USA
Bureau of Accreditation (BoA), Vietnam
Combined standard uncertainty

Standard uncertainty of the result of a measurement when that result is obtained from the values of a
number of other quantities, equal to the positive square root of a sum of terms, the terms being the
variances or covariances of these other quantities weighed according to how the measurement result
varies with changes in these quantities.

Ref: RAL110BEN
Appendices
Glossary of Terms
Contaminant (Effect)
Undesirable effect, resulting from contamination. Most commonly, this is the effect exerted by a
serum on that which follows or precedes it. It may also arise from contaminating effects between
reagents.
Control
Substance used for monitoring the performance of an analytical process or instrument.
Conventionally true value (of a quantity)

Value attributed to a specific and recognized quantity, sometimes by convention, as the
representative for an appropriate uncertainty for a given use.
Correction
Value that is algebraically added to the raw result of a measurement to compensate for a systematic
error.
■ the correction is equal to the opposite of the estimated systematic error
■ since the systematic error cannot be precisely known, the compensation cannot be complete.
Correlation coefficient
Quotient of the covariance of two characteristics by the product of their standard-deviations. It
expresses the possible relationship between two variables that are known to be independent. Its value
must only be tested in comparison with zero according to a chosen risk. It is usually of no interest in
technical comparisons.
Coverage factor
Numerical factor used as a multiplier of the combined standard uncertainty to obtain the expanded
uncertainty.
Default setting
Original factory setting.
Detection limit (XP T 90-210)

The smallest quantity of an analyte to be examined in a sample that can be detected and considered
as being different from the value of the blank (with a given probability), but not necessarily quantified.
Two risks need to be taken into account:
■ the risk of considering the substance present in the sample when in fact its quantity is nil.
■ the risk of considering a substance absent when in fact its quantity is not nil.
Deviation
Value minus its reference value.
Drift
Slow variation over time of a metrological characteristic of a measuring instrument.
Error
Result of a measurement minus a true value of the measurand (Bias).

Ref: RAL110BEN
Appendices
Glossary of Terms
Error of trueness (of a measuring instrument)

Systematic indication error of a measuring instrument. The trueness error is normally estimated by
taking into account the mean of the indication error on an appropriate number of repeated
observations.
Evaluation of type A uncertainty

Method of evaluating the uncertainty through the statistical analysis of a series of observations.
Evaluation of type B uncertainty

Method of evaluation using methods other than the statistical analysis of a series of observations.
Exactitude (Precision)
Closeness of the agreement between the result of a measurement and the true value of the
measurand.
Expanded uncertainty
Quantity defining an interval about the result of a measurement that may be expected to encompass a
large fraction of the distribution of values that could reasonably be attributed to the measurand.
■ The fraction may be viewed as the probability or level of confidence of the interval.
■ The association of a specific level of confidence with the interval defined by the expanded
uncertainty requires explicit or implicit assumptions regarding the probability distribution
characterized by the measurement result and its combined standard uncertainty. The level of
confidence that may be attributed to this interval can be known only to the extent to which such
assumptions may be justified.
■ The expanded uncertainty is sometimes known as the overall uncertainty.
Femtoliter (fL)
One quadrillionth (10-15) of a liter.
Grading
Material positioning of each marker (possibly of certain principal markers only) of a measurement
instrument according to the value of the measurand.
Linearity (XP T 90-210)

Capacity of a method of analysis, within a certain interval, to provide a value of information or results
proportional to the quantity of analyte to be assayed in the laboratory sample. This proportionality is
expressed using a previously defined mathematical expression. The limits of linearity are the
experimental limits of quantities between which a linear standard model can be applied with a known
level of confidence (generally taken as being equal to 1%).
L.I.S.
Laboratory Information System
Lot number
Manufacturer’s code that identifies a batch of product (either reagents, controls or calibrators).
Matrix
Environment in which the analyte is found.

Ref: RAL110BEN
Appendices
Glossary of Terms
Mean, m
The sum of observations divided by their number. Unless otherwise indicated, the term “mean”
designates the arithmetic value.
Measurand
Specific quantity subjected to measurement.
Measurement
A series of operations whose aim is to determine a value of a quantity.
Noise
Corresponds to random variations of the measurement signal for a given level. It is measured by the
standard deviation of a series of at least 30 measurements of the signal, at the level in question.
Operating range
Range of results over which the instrument displays, prints and transmits data.
Parameter
Component of blood that the instrument measures and reports.
Performance criteria
Parameters characterizing the analytical procedure (linearity, repeatability, trueness, etc.)
Platelet concentrate
Labile blood product, composed of platelets, produced by blood bank centers and intended for
transfusion.
PRP (Platelet Rich Plasma)

Cellular suspension in the plasma, high platelet concentration obtained by sedimentation from a
whole blood sample.
Quality control (QC)

Comprehensive set of procedures that a laboratory establishes to ensure that the instrument is
working accurately and precisely.
Quantitation limit (XP T 90-210)

The smallest quantity of an analyte to be analyzed in a sample that can be determined quantitatively
under the experimental conditions described in the method with a defined variability (determined
coefficient of variation).
Random error
Result of a measurement minus the mean of an infinite number of measurements of the same
measurand, undertaken under conditions of repeatability.
■ the random error is equal to the error minus the systematic error.
■ since one can only perform a finite number of measurements, it is only possible to determine an
estimation of the random error.

Ref: RAL110BEN
Appendices
Glossary of Terms
Range of measurement (field of analysis, scope of measurement)

Set of values of the measurand for which the error of a measuring instrument is presumed to be
comprised between specified limits.
Reagent blank
Corresponds to the signal resulting from the reagent(s) used during an assay or a measurement of
catalytic activity. The sample is replaced by an equal volume of an appropriate solvent.
Reference material (Calibrator, reference values)

Material or substance of which one (or several) value(s) of the property(ies) is (are) sufficiently
homogeneous and well-defined to enable it to be used to calibrate a piece of equipment, evaluate a
measuring method, or attribute values to materials.
Reference technique (reference method)

Internationally recognized technique whose accuracy has been evaluated in comparison with a
definitive technique or following a complete and detailed study. The principle, reagents, apparatus,
operating procedure, and the calculations are precisely defined.
Reference values
Results obtained for a given component in a reference population whose individuals are exempt from
disease or treatments that may alter their values. The reference values may vary, notably according to
the geographic origin, sex, and age of individuals. They are usually expressed as a function of lower
and upper limits that have been determined via statistical studies. They may be established by the
biologist, according to the analytical techniques used, or possibly verified when data from scientific
publications is used. The expression «reference value» is preferable to «usual value» or «normal
value».
Reliability (Precision)
Aptitude of a measuring instrument to give very similar indications during the repeated application of
the same measurand under the same measurement conditions.
Repeatability
Closeness of the agreement between the results of successive measurements of the same
measurand, measurements undertaken entirely in the same conditions of measurement.
Reproducibilty
Closeness of the agreement between the results of measurements of the same measurand,
measurements undertaken under a variety of measurement conditions.
Result of a measurement
Value attributed to a measurand, obtained by measurement.
Robustness
The “robustness” of an analysis technique implies its capacity to be unaffected by small, but
deliberate variations in parameters of the method, and it gives an indication about its reliability under
normal conditions of use.
Sensitivity (diagnostic)
The probability that a device gives a positive result in the presence of a target marker.

Ref: RAL110BEN
Appendices
Glossary of Terms
Sensitivity of a technique
Relationship between the variation of the measured signal to the concentration unit of the analyte
being studied.
Shutdown cycle
Cleans the instrument’s fluidic lines and apertures to help prevent residue build-up.
Specificity (diagnostic)
The probability that a device gives a negative result in the absence of the target marker.
Specificity (XP T 90-210)

Property of an analysis method to be exclusively suited to the determination of the quantity of the
analyte under consideration.
Specimen
To avoid any confusion with the term sample (in the following context: group of individuals from a
population), it is preferable to use the term specimen to designate a biological sample (blood
specimen, urine specimen, etc.)
Specimen blank
Signal resulting from certain properties of the environment in which the analyte is found. Ideally, it
results from a signal measurement undertaken under the conditions of the reaction on the sample that
does not contain the analyte, or on the sample following elimination or inactivation of the analyte.
Standard
Materialized measurement, measuring apparatus, reference material or measurement system
designed to define, undertake, store, or reproduce a unit or one or several values of a quantity to
serve as a reference.
Standard Deviation (SD)

Measure of variation within a group samples or within a population.
Standard Error of the Mean (SEM)

Statistical parameter indicating the dispersion of values at the level of the mean of a series of
measurements.
Standard uncertainty
Uncertainty of the result of a measurement expressed as a standard deviation.
Startup cycle
Ensures that the instrument is ready to run; includes performing a background test.
State of the art, (Performance of the)

Designates the quality of currently available analytical methods. This is usually assessed on the basis
of inter-laboratory surveys.
Systematic error
Mean that would be obtained from an infinite number of measurements of a same measurand,
undertaken under the conditions of repeatability, minus a true value of the measurand.

Ref: RAL110BEN
Appendices
Glossary of Terms
■ the systematic error is equal to the error minus the random error.
■ as with the true value, the systematic error and its causes cannot be fully known.
■ for a measuring instrument, see "trueness error".
Traceability (VIM)
Property of the result of a measurement or a standard such that it can be linked to determined
references, generally national or international standards, via the intermediary of an uninterrupted chain
of comparisons, all with determined uncertainties.
■ This concept is often expressed by the adjective «traceable».
■ The uninterrupted chain of comparisons is known as the chain of standardization or chain of
linkage of standards.
■ The manner in which the linkage to the standards is performed is known as the link to standards.
Traceability (ISO 9000 : 2000): ability to find the history, implementation, or location of that which
is being examined.
Trueness
Aptitude of a measuring instrument to give results that are exempt from systematic error.
Uncertainty
Parameter associated with the result of a measurand that characterizes the dispersion of values that
could reasonably by attributed to the measurand.
Validation (analytical and biological)

This is the set of procedures used to ensure that a technique has the required reliability to meet the
quality control requirements in the state of the art. The validation generally comprises two stages: a
technical validation and a biological validation. The first consists, following a series of assays, of
verifying with appropriate controls that the principal errors have been maintained within acceptable
limits. The second involves ensuring the coherence of the result in its clinical context, by comparing it
with any previous results and with the results of other analyses requested for exploring the same
function.
Validation (validation of methods)

Verification process that involves comparing the values of performance criteria, as determined during
the characterization study or experimentation phase (test phase) of the analytical method, to those
initially expected or assigned (acceptable limits, objectives to be attained), and then to declare
whether the method of analysis is valid or not (see definition of the standard EN ISO/CEI 17025,
§5.4.5.1).
Validation technique
A validation technique is a technique that, following exhaustive study (literary and experimental
research), is designated for use as a reference of exactitude, for comparative assays or titrations of
control sera. It is chosen from amongst the reference techniques or selected by national or
international companies, or by consensus. It has acceptable and recognized levels of precision and
practicability. It should be described in detail, tested in several laboratories, and include the definition
of the equipment, controls required on the apparatus, the description of the various reagents and
specifications, the conditions of storage and use of the reagents, the operating procedure, the
calibration or standardization method, the nature of the calibrators, and the control method.
Verification (EN ISO 10012)

Confirmation by examination and establishment of proofs that the specified requirements have been
met.

Ref: RAL110BEN
Appendices
Glossary of Terms
Whole blood
Non-diluted blood (blood and anticoagulant only).

Ref: RAL110BEN
Appendices
References
2. References
2.1. Regulatory References
■ Arrêté du 26 novembre 1999 relatif à la bonne exécution des analyses de biologie médicale.
(GBEA) J.O. Numéro 287 du 11 décembre 1999, page 18441 (NOR : MESP9923609A)
■ Directive 98/79/CE du Parlement européen et du Conseil, du 27 octobre 1998, relative aux
dispositifs médicaux de diagnostic in vitro (http://www.lne.fr/publications/directives/98-79.pdf)
■ Décret n° 2004-108 du 4 février 2004 relatif aux dispositifs médicaux de diagnostic in vitro et
modifiant le code de la santé publique (deuxième partie : Décrets en Conseil d'Etat): J.O. du 6
février 2004, page 2577, NOR: SANP0324626D
■ Essential Criteria for Quality Systems of Medical Laboratories (Eur J Clin Chem Clin Biochem
1997; 35(2): 123-132 © 1997) Walter de Gruyter Berlin New York
■ CLIA (Clinical Laboratory Improvement Amendments) regulation (http://www.westgard.com/
cliafinalrule4.htm)
2.2. General Standardization References
■ EDMA Position paper : Laboratory Accreditation (http://www.edma-ivd.be/fileadmin/

upl_documents/Position_Papers/Accrediation_Position_Paper_FINAL.pdf)
■ EDMA Position paper : Estimation of uncertainty of measurement in medical laboratories (http://
www.edma-ivd.be/fileadmin/upl_documents/Position_Papers/Estimation_of_uncertainty.pdf)
■ Laboratoires d'analyses de biologie médicale - Exigences particulières concernant la qualité et la
compétence. NF EN ISO 15189: 2007 (AFNOR).
■ Normes fondamentales - Vocabulaire des termes fondamentaux et généraux de métrologie (VIM).
NF X 07-001: Décembre 1994 (AFNOR).
■ Statistique - Vocabulaire et symboles - Partie 1 : probabilité et termes statistiques généraux. NF
ISO 3534-1: Décembre 1993 (AFNOR).
■ Qualité de l'eau - Protocole d'évaluation initiale des performances d'une méthode dans un
laboratoire. XP T90-210 : Septembre 2008 (AFNOR)
■ Métrologie - Systèmes de management de la mesure - Exigences pour les processus et les
équipements de mesure. NF EN ISO 10012 : Septembre 2003 (AFNOR).
■ Guide pour l'expression de l'incertitude de mesure (GUM). NF ENV 13005: Août 1999 (AFNOR).
■ Application de la statistique - Exactitude (justesse et fidélité) des résultats et méthodes de mesure
- Parties 1-6. NF ISO 5725: Décembre1994 et rectificatifs techniques (AFNOR).
■ Normes fondamentales - Métrologie et applications de la statistique - Aide à la démarche pour
l'estimation et l'utilisation de l'incertitude des mesures et des résultats d'essais. FD X07- 021:
Octobre 1999 (AFNOR).
■ Estimer l'incertitude, Mesures et Essai. C. Perruchet et M. Priel, 2000 (AFNOR).
■ Lignes directrices relatives à l'utilisation d'estimations de la répétabilité, de la reproductibilité et de
justesse dans l'évaluation de l'incertitude de mesure. ISO/TS 21748: Mars 2004 (AFNOR).
■ Systèmes de management de la qualité - Principes essentiels et vocabulaire. NF EN ISO 9000 :
Décembre 2000 (AFNOR).

Ref: RAL110BEN
Appendices
References
2.3. COFRAC-EA Documentation
■ Documents COFRAC d'accréditation COFRAC en Biologie Médicale (Programmes n° 143,145,

respectivement, Analyses en Biochimie, Analyses en Hématologie).
■ Document COFRAC LAB LABM REF 04, "Accréditation des Laboratoires d'Analyses de Biologie
Médicale suivant la norme ISO 15189 - Politique COFRAC".
■ Document COFRAC LAB Inf 09 (1133), "Procédure de validation interne des méthodes d'essais".
■ Document EA-4/16 "Lignes directrices d'EA pour l'expression de l'incertitude des résultats
d'essais quantitatifs" (http://www.lne.fr/publications/EA_4_16_anglais_francais.pdf).
■ Document COFRAC LAB GTA 04, Guide de validation des méthodes en biologie médicale.
■ Document COFRAC LAB GTA 06, Les contrôles de la qualité analytique en biologie médicale.
■ Document COFRAC LAB GTA 14, Guide d'évaluation des incertitudes de mesures des analyses
de biologie médicale.
2.4. Method Validation
■ The Fitness for Purpose of Analytical Methods: A Laboratory Guide to Method Validation and
Related Topics. Eurachem Guides (1998).
■ A WHO guide to good manufacturing practice requirements - Part 2: Validation. World Health
Organisation (1997).
■ Analyses de biologie médicale : spécification et normes d'acceptabilité à l'usage de la validation
des techniques. A. Vassault, D. Grafmeyer, J. de Graeve, R. Cohen, A. Beaudonnet, J. Bienvenu.
Ann Biol Clin 1999, 57 : 685-95.
■ AIDE MEMOIRE D’HEMATOLOGIE, 1998. Prof. C. Sultan / M. Gouault - Helman / M. Imbert,
Service Central d’Hématologie de l’Hôpital Henri Mondor, Faculté de médecine de Créteil (Paris
XII).
■ Protocole de validation de techniques (Protocole Valtec) - Document B. Ann. Biol. Clin 44, 686-
745. Vassault A., Grafmeyer D., Naudin C., Dumont G., Bailly M., Henny J., Gerhardt MF., Georges
P. et les membres de la commission de Validation de techniques de la SFBC (1986).
■ Dictionnaire des termes à l'usage de la validation des techniques, Commission validation de
technique SFBC, Ann Biol Clin 1986, 44: 679-85.
■ Guidelines for the Evaluation of blood cell analyzers including those used for differential leukocyte
and reticulocyte counting and cell marker applications. International Council for Standardization in
Hematology; Clin. Lab. Haemat. 1994, 16, 157-174.
■ Guidelines for the evaluation of analyzers in clinical chemistry ECCLS Document 3 n°2. Haeckel R,
Busch EW, Jennings RD, Truchaud A (1986) Beuth Verlag Berlin.
■ Proposed quality specifications for the acceptability of analytical systems for clinical chemistry.
Fraser CG, Hyltoft Petersen P, Ricos C, Haeckel R. Eur J Clin Chem Clin Biochem 1992, 30 : 311-
317.
■ Evaluation de l'accord entre trois automates d'hématologie. F. Hennouchi, S. Anselme Martin, V.
Chanteperdrix, N. Roux Bouisson, B. Polack, P. Mossuz. Ann Biol Clin 2002, 60 : 351-5.
■ Lecture critique de l'hémogramme : valeurs seuils à reconnaître comme probablement
pathologiques et principales variations non pathologiques – ANAES – http://www.has-sante.fr/
portail/jcms/c_271914
■ Age-based Reference Intervals from data is the third National Health and Nutrition Examination
Survey (NHANES III) - http://www.dgrhoads.com/refint/index.htm
■ DESIRABLE SPECIFICATIONS FOR TOTAL ERROR, IMPRECISION, AND BIAS, DERIVED FROM
BIOLOGIC VARIATION by Ricos C, Alvarez V, Cava F, Garcia-Lario JV, Hernandez A, Jimenez CV,
Minchinela J, Perich C, Simon M. "Current databases on biologic variation: pros, cons and
progress." Scand J Clin Lab Invest 1999;59:491-500 (mis à jour en 2008) - http://
www.westgard.com/biodatabase1.htm
■ Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory; Approved
Guideline ; CLSI Document C28-A3 ((ISBN 1-56238-682-4)), 2008.

Ref: RAL110BEN
Appendices
References
■ Assessment of the Clinical Accuracy of Laboratory Tests Using Receiver Operating Characteristic
(ROC) Plots; Approved Guideline, CLSI document GP10 (ISBN 1-56238-285-3) 1995.
■ Calibration and Quality Control of Automated Hematology Analyzers; CLSI Document H38-P (ISBN
1-56238-398-1), 1999.
■ Evaluation of Precision Performance of Clinical Chemistry Devices; Approved Guideline, CLSI
document EP5-A2 (ISBN 1-56238-542-9) 2004.
■ Validation, Verification, and Quality Assurance of Automated Hematology Anaysers; Approved
Standard ; CLSI Document H26-A2 (ISBN 1-56238-728-6), 2010.
■ Protocols for Determination of Limits of Detection and Limits of Quantification; Approved
Guideline, CLSI document EP17-A I(SBN 1-56238-551-8) 2004.
■ Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline ; CLSI
Document EP9-A2 ((ISBN 1-56238-472-4)), 2002.
■ Reference Leukocyte (WBC) Differential count (Proportional) and Evaluation of Instrumental
Methods; Approved Guideline ; CLSI Document H20-A2 ((ISBN 1-56238-628-X)), 2007.
■ Procedures for the Handling and Processing of Blood Specimens; Approved Guideline; NCCLS
Document H18-A4 (ISBN 1-56238-724-3), 2010.
■ User protocol for Evaluation of Qualitative Test Performance; Approved Guideline - Second
Edition, Approved Guideline ; CLSI Document EP12-A2 ((ISBN 1-56238-654-9)), 2008.
■ Evaluation of the Linearity of Quantitative measurement procedures: a statistical Approach;
Approved Guideline, NCCLS document EP6-A I(SBN 1-56238-498-8) 2003.
■ Interference Testing in Clinical Chemistry: Approved Guideline ; NCCLS Document EP7-A2 ((ISBN
1-56238-584-4)), 2005.
■ ICSH guidance (Clin. Lab. Haemat.1994, 16, 157-174).
■ H20-A2 : Reference Leukocyte (WBC) Differential Count (Proportional) and Evaluation of
Instrument Methods, January 2007.
2.5. Websites
■ James O. Westgard, PhD, www.westgard.com

■ Ph. Marquis, www.multiqc.com
■ Cofrac, Comité Français d'Accréditation, www.cofrac.fr
■ AFNOR, Association Française de Normalisation, www.afnor.fr
■ EA, European co-operation for Accréditation, www.european-accreditation.org
■ IAF- International Accreditation Forum, www.iaf.nu
■ ILAC- International Laboratory Accreditation Cooperation, www.ilac.org
■ ILAC News, http://www.ilac.org/news.html
■ ISO- International Organization for Standardization, www.iso.org
■ PROBIOQUAL - Association pour la promotion du Contrôle de Qualité en Biologie,
www.probioqual.com
■ EC4: European Communities Confederation of Clinical Chemistry, www.ec-4.org

Ref: RAL110BEN
Index
A I
Accuracy, 48 Individual Machine Data Sheets, 55
Agglutinated red blood cells, 42,43,44 Instructions for Use, 55
Analysis Data, 56 Interfering substances, 45
Analytical measuring interval, 37 GRA, 45
Analytical Procedures, 58 HCT, 43
Analytical Sensitivity, 36 HGB, 42
Approach to Trueness, 30 LYM, 44
MCH, 43
B MCHC, 43
Bias calculation, 30 MCV, 43
Biological risk, 24 MON, 44
Blood transfusion, 43 MPV, 44
PLT, 43
C RBC, 42
Calibration, 26,59 RDW, 43
Certificates, 16 WBC, 41
Chemical risk, 24 ISO standards, 16
Chemotherapy, 41,43,44
Citrate blood, 43 L
Cold agglutinins, 42 Labeling, 54
Comparison with a reference instrument, Laboratory Equipment, 26
48 Laboratory Premises, 23
Contamination, 38 Large platelets, 43
Continuous Improvement, 12 Leukemia, 41
Copyright, 6 Linearity, 37
Correlation, 48 Lipids, 42
Cryoglobulins, 41 Logos
Definition, 6
D
Declaration of Conformity, 5 M
Disposal Macro platelets, 43,44
Biological Materials, 56 Macrothrombocytes, 41
Chemical Products, 56 Maintenance, 26
Multiple myeloma, 41
E
Energy Consumption, 26 N
Environmental Conditions, 23 Notice of Liability, 5
Environmental protection, 24 Nutritional deficiency, 43
Erythroblasts, 41
External Services, 11 O
Operating Conditions, 23
F
Field of Analysis, 36 P
Parameters, 36
H CBC Parameters, 27
Hemolysis, 41,43 Performance, 27
Personnel, 22
Pictograms

Ref: RAL110BEN
Definition, 6
Platelet agglutination, 41,43
Pre-analytical Procedures, 57
Processing Documents, 9
Q
Q.H.S.E. policy, 16
QCP, 32
Quality Management System, 8
Quality of Analytical Procedures, 59
Quality Records, 13
R
RBC
Inclusions, 43
Red blood cells agglutination, 43
Reference Values, 46
References
COFRAC-EA Documentation, 79
General Standardization, 78
Regulatory, 78
Websites, 80
Repeatability, 29
Reproducibility, 28
Results Report, 65
Review of Contracts, 10
Robustness, 39
S
Sigma Calculation, 34
Small erythrocytes, 44
Specimen for Sampling, 57
Stability, 39
Storage Conditions, 23
Supplies, 11
Symbols definition
Caution, 6
Danger, 6
Nota, 6
T
Technical Records, 13
Technical Requirements, 0
Turbidity, 42
U
Uncertainty Calculation, 32
Unlysed RBC, 41
Use of Computers, 56
W
Waste Disposal, 27
Westgard, 34

Ref: RAL110BEN

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Hematology Analyzer

Performance and Reference:

Performance and Reference: Tools for Accreditation

Requirements Relating to Management...................................................................7

4. External Services and Supplies.......................................................................................11

6. Quality Records and Technical Records.....................................................................13

2. Laboratory Premises and Environmental Conditions............................................23

6. Ensuring Quality of Examination Procedures...........................................................59

Performance and Reference: Tools for Accreditation i

Performance and Reference: Tools for Accreditation 3

Index Reference Software Version Date

4 Performance and Reference: Tools for Accreditation

2.1. Declaration of Conformity

2.2. Notice of Liability

Linux is a registered trademark of Linus Torvalds.

Performance and Reference: Tools for Accreditation 5

2.5. Document Symbols

2.6. Copyright ® 2011 by HORIBA ABX SAS

HORIBA ABX SAS

2.7. Document Intended Use

6 Performance and Reference: Tools for Accreditation

1. Quality Management System.............................................................................................8

4. External Services and Supplies.......................................................................................11

6. Quality Records and Technical Records.....................................................................13

Performance and Reference: Tools for Accreditation 7

1. Quality Management System

■ § 4.2.5 of the standard ISO 15189 : 2007

8 Performance and Reference: Tools for Accreditation

■ § 4.3.1 of the standard ISO 15189 : 2007

Performance and Reference: Tools for Accreditation 9

■ § 4.4 of the standard ISO 15189 : 2007

10 Performance and Reference: Tools for Accreditation

4. External Services and Supplies

■ § 4.6.1 of the standard ISO 15189 : 2007

■ § 4.6.3 of the standard ISO 15189 : 2007

■ § 4.6.4 of the standard ISO 15189 : 2007

Performance and Reference: Tools for Accreditation 11

■ § 4.12.5 of the standard ISO 15189 : 2007

12 Performance and Reference: Tools for Accreditation

6. Quality Records and Technical Records

■ § 4.13.3 of the standard ISO 15189 : 2007

National, regional and local regulations may apply.

■ § 4.15.2 of the standard ISO 15189 : 2007

Performance and Reference: Tools for Accreditation 13

Identification of the Company

Legal type of company: SAS (Société par Actions Simplifiée)

International Marketing Director Alfredo Braga

International Technical Service Manager Jean-Louis Taravella

Quality & Regulatory Affairs Director Arnaud Pradel

Reacto-Vigilance Manager Arnaud Pradel

Main Activities sectors

14 Performance and Reference: Tools for Accreditation

Policy related to Quality

Performance and Reference: Tools for Accreditation 15

16 Performance and Reference: Tools for Accreditation

Performance and Reference: Tools for Accreditation 17

18 Performance and Reference: Tools for Accreditation

Performance and Reference: Tools for Accreditation 19

20 Performance and Reference: Tools for Accreditation

2. Laboratory Premises and Environmental Conditions............................................23

6. Ensuring Quality of Examination Procedures...........................................................59

Performance and Reference: Tools for Accreditation 21

■ § 5.1.2.d of the standard ISO 15189 : 2007

22 Performance and Reference: Tools for Accreditation

_____ TRUENESS _ SYSTEMATIC _______ BIAS

_____ _ TOTAL _______ UNCERTAINTY

_____ PRECISION _ RANDOM _______ STANDARD