conversion/tmp/activity - Task - Scratch/582676692.docx: Document Issued: 12/6/17 Page 1 of 14

A2LA
Document Issued:
C231 - Specific Checklist: Threat Agent Testing Laboratory Program Testing 12/6/17
Page 1 of 14
The following pages present the A2LA R231 – Specific Requirements: Threat Agent Testing Laboratory Accreditation Program. All laboratories participating
in this program shall be assessed to ISO/IEC 17025:2005 and to these requirements. Management system documentation and supporting records must be
available for the assessor’s review.
Laboratory Instructions: This checklist will be completed by the assessor as part of their onsite assessment. Therefore, it will not be required to be submitted as
part of the laboratory’s application for accreditation.
A2LA Assessor Instructions: Review the laboratory’s documented management system to verify compliance with the applicable requirements. Assess
to verify that the documented management system is indeed implemented as described. Place a tick mark in the yes (Y), no (N), or not applicable (NA)
space for each checklist item. Please note that for all N/A indications, you must document the reason why this requirement is N/A in the comments
section. Record comments related to any requirement in the space provided. Record comments related to tests on separate sheets and/or on the Test
Method Matrix. All deficiencies must be identified and explained in the assessor deficiency report. Assess the laboratory’s technical competence to
perform specific tests or specific types of tests. Please also complete the separate C104 – General Checklist: Requirements When Making Reference to
A2LA Accredited Status, C113 – Specific Checklist: A2LA Policy on Measurement Traceability for Life Sciences Testing Laboratories , and C106 – General
Checklist: Proficiency Testing for ISO/IEC 17025 Laboratories checklists. The laboratories themselves are not required to complete C104-C106 or C113
prior to the assessment.
IMPORTANT NOTE: An asterisk (*) in the comments section indicates that the assessor must document the specific traceable objective evidence reviewed in
association with that requirement. Objective evidence information is mandatory for those clauses.
© 2017 by A2LA.
All rights reserved. No part of this document may be reproduced in any form or by any means without the prior written permission of A2LA.
/conversion/tmp/activity_task_scratch/582676692.docx
A2LA
Document Issued:
Page 2 of 14
To the best of my knowledge, all laboratory document references below, as well as actual practice, have been assessed for compliance to the relevant clauses of
this document. I hereby attest that all ‘Yes’ marked compliance clauses, whether initialed or not, meet the aforementioned requirements. Any areas of
noncompliance have been fully described in the Assessor Deficiency Report.
Master Code: Assessment ID:

Assessor: Assessor Signature &
Date:
A2LA
Document Issued:
Page 3 of 14
{RESERVED FOR A2LA ASSESSORS ONLY}
Requirement Reference Compliance

Comments
Y N NA
4.0 Management Requirements

4.13 Control of Records
4.13.2.1: Records of original observations, derived data and
sufficient information to establish an audit trail are required in order
to enable the test to be repeated under conditions as close as
possible to the original. These records include, but may not be
limited to:
Incubator temperature (start/end or at least twice daily);
Oven temperature (start/end or at least twice daily);
Time incubation started (if no timer set);
Time incubation ended (if no timer set);
Extraction time started (if no timer set);
Extraction time ended (if no timer set);
Initials of analyst involved in each step of the preparation and
testing process;
Lot numbers of reagents used (commercial and prepared);
Lot numbers of media used (commercial and prepared);
Reagent & media preparation records (including lot numbers of all
components);
Lot numbers of materials critical to the process (ex. extraction
filters, petri dishes, etc.);
Lot numbers and Certificates of Analysis of Certified Reference
Materials (CRMs) and Reference Materials (RMs) (also see
5.6.3.2);
A2LA
Document Issued:
Page 4 of 14

Comments
Y N NA
Identification of equipment used that is critical to the process (ex.

pipettors, ovens, incubators, HPLC instruments, balances, etc.);
Purchasing records, including POs and vendor evaluation/selection
records;
QC of materials received critical to the test process.
5.0 Technical Requirements
5.3 Accommodation and Environmental Conditions
5.3.2 The laboratory shall monitor, control and record *
environmental conditions as required by the relevant specifications,
methods and procedures or where they influence the quality of the
results. This includes the following:
Air exposure plates - daily (when plating is performed);
Environmental swabs (contamination control swabs)
of the test area to ensure cross-contamination has not occurred -
daily (when testing performed)– these swabs may be tested with
daily samples, but are required to be tested if suspect target is
identified;
Restricted access to the test area when testing is in process,
restricted access of unauthorized personnel, procedures to handle
breach in access;
Evaluation of previous use of the area before new testing activities
commence.
5.4 Test Methods and Method Validation
A2LA
Document Issued:
Page 5 of 14

Comments
Y N NA
5.4.5.1 Prior to developing the proposed methods:

5.4.5.1.1 The intended use of the method shall be defined;
5.4.5.1.2 The intended use should be aligned with current
capabilities;
5.4.5.1.3 A written protocol which includes all of the following
elements shall be prepared:
Intended use and criteria for use;
Assay principle and safety precautions;
Acceptable sample types, sample collection method, preparation,
preservation, storage and transportation conditions;
Description of reagents supplied or directions for preparation and
QC;
Description of quality controls to be used;
Detailed instructions on how to perform the method;
Detailed instructions on how to interpret and report the result;
Any limitations of the method that are known or suspected;
A summary of the performance characteristics of the method;
A statement on the target uncertainty of measurements for each
analyte (analytical goal for accuracy) in order for the values to be fit
for their intended purpose.
5.4.5.2 Methods shall be validated whenever any of the following
occur:
5.4.5.2.1 Development of a proposed new official method;
5.4.5.2.2 Expansion of the scope of an existing method to include
additional analytes or new matrices;
A2LA
Document Issued:
Page 6 of 14

Comments
Y N NA
5.4.5.2.3 Modification of a method’s range beyond validated levels;

5.4.5.2.4 Modification of a method that may alter its performance *
specifications. All but the most trivial of changes shall be evaluated
for effects on method performance. This includes:
Changes to the fundamental science of an existing method;
Equivalence issues such as substitutions of reagents/apparatus, or
changes to some instrumental parameters since it is difficult to
predict the results of any change.
5.4.5.3 Performance specifications required to validate a method
include the following:
5.4.5.3.1 Performance specifications that should be determined to
validate a method will vary depending on the intended use, the type
of method being validated, and the degree to which it has previously
been validated. All methods submitted shall have all the proper
controls and the parameters for calibrating and operating the method
instrumentation included in the written procedure.
5.4.5.3.2 Typical validation characteristics which shall be
considered are the following:
5.4.5.3.2.1 Characteristics of quantitative methods:
Method uncertainty at range of use;
Minimum quantifiable concentration;
Applicable analyte concentration range;
Accuracy (trueness);
Precision (repeatability);
Analytical specificity;
Linearity;
A2LA
Document Issued:
Page 7 of 14

Comments
Y N NA
Ruggedness/robustness
5.4.5.3.2.2 Characteristic for qualitative methods:
Evaluate for reliable identification of an analyte at or below some
target level;
Sensitivity;
Specificity;
Limit of detection (See Detection limit, MDL, MDC);
Ruggedness/robustness.
5.4.5.3.3: The following validation tools shall be used to
demonstrate the ability to meet method specifications of
performance:
Blanks: Use of various types of blanks enables assessment of how
much of the result is attributable to the analyte in relation to other
causes;
Reference materials and certified reference materials with the
typical interferences expected. Use of known materials can be
incorporated to assess the accuracy of the method, as well as for
obtaining information on interferences;
Fortified (spiked) materials and solutions: Recovery determinations
can be estimated from fortification or spiking with a known amount
of analyte. (Note: Understand that spiked recovery may not be
truly representative of recovery from naturally incurred analytes);
Repeatability: Replicate analyses provide a means of checking for
changes in precision in an analytical process which could adversely
affect the results;
A2LA
Document Issued:
Page 8 of 14

Comments
Y N NA
Statistics: Statistical techniques are employed to evaluate accuracy,

trueness and precision, linear range, limits of detection and
quantification, and measurement uncertainty.
5.4.5.3.4 General Validation Protocol
5.4.5.3.4.1 The following provides practices that shall be used to
determine method performance characteristics:
Quantitative measurements, e.g. determine limit of quantification
(LOQ), linear response. Minimally need LOD;
Prepare and analyze spiked blanks, matrix samples of known *
concentration utilizing one to three different concentration levels:
low, medium, high based on the intended use of the method. These
samples are carried through the complete sample preparation
procedure, extraction and analytical steps of a particular method.
Matrix effects shall be assessed with these samples. Accuracy or
bias and precision are calculated from these results; data will also
evaluate robustness of the method resulting from changes in the
sample matrix. (Note: Proper certified reference materials and
reference standards are used when available.);
Assure that adequate sample replicates are performed and that
results from replicate measurements of each analyte are compared;
Analyze blanks (reagent and matrix) and compare these results to
the reported limit of detection;
Evaluate interferences: spectral, physical, chemical or memory by
analyzing samples containing various suspected interferences in the
presence of the measurand.
5.4.5.3.5 Validation of Methods (Original, New or Modified)
A2LA
Document Issued:
Page 9 of 14

Comments
Y N NA
5.4.5.3.5.1 The following practices shall include, but not be limited

to, matrix extensions and platform changes:
In cases where the sample preparation and/or the extraction
procedure/analytical method is modified from the existing test
procedure and protocol, the new method shall demonstrate that the
modifications do not adversely affect the precision and accuracy or
bias of the data obtained;
In order to implement the modified method, the standard or existing
method is first performed. The modified method is then verified
against the original method validation protocol;
For original or new methods the authors shall pick a validation level
that is suitable for their situation;
Statistical methods are employed to verify performance between the
original validated and new method sample means and to determine
the degree of accuracy. (Note, for example: The t-test assesses
whether the means of two groups are statistically different. The t-
test is to be less than or equal to the t-critical value. The F-test is
used to determine the significance of difference between two sample
variances. The F value is to be less than or equal to the F-critical
value). Note: See Tables for details.
5.4.6: Estimation of Uncertainty of Measurement
5.4.6.2 Measurement Uncertainty is required to be estimated for
tests with a qualitative reported result, if the result is based on
quantitative endpoint, such as presence of a compound above a
threshold value.
5.6 Measurement Traceability
A2LA
Document Issued:
Page 10 of 14

Comments
Y N NA
5.6.3.2 In the event that CRMs are not available from an Asia *
Pacific Laboratory Accreditation Cooperation (APLAC) recognized,
ISO/IEC 17034accredited source, the laboratory shall provide
records of material evaluation and traceability consistent with
A2LA Policy P102a, “Policy on Reference Material Traceability for
Life Sciences Testing Laboratories”. This includes the following
records:
5.6.3.2.1. When Reference Materials are used as process
controls:
Unique identifier or lot number of the material (required);
Description including (as appropriate):
Origin, (e.g. inorganic, human or animal, vegetable, or microbial);
Who and where manufactured (may be generated in-house or
provided by supplier);
Other pertinent information;
Records of use and process control results (required). *
5.6.3.2.2 When Reference Materials are used to validate
methods:
Unique identifier or lot number (required);
Origin (if applicable) (e.g. inorganic, human or animal, vegetable,
or microbial);
Issuing authority (if applicable) (e.g. WHO, BCR, IRMM, USP,
ATCC);
Records of characterization (if applicable); *
A2LA
Document Issued:
Page 11 of 14

Comments
Y N NA
Molecular form(s) of or surrogate for the analyte (e.g. steric isomer

for an amino acid, or glycerol for glycerol ester);
Matrix (e.g. buffered bovine albumin solution);
State(s) of aggregation (gas, liquid, solid);
Phase(s) (solution, suspension, lyophilized);
Records of use (required); *
Records of Materials validation activities including an appropriate *
validation plan, acceptance criteria, and results (as appropriate).
5.6.3.2.3 When Reference Materials used are obtained from an authoritative source
or prepared by a published standard procedure (to demonstrate operational traceability):
Unique identifier or lot number (required);
Origin (if applicable) (e.g. inorganic, human or animal, vegetable,
or microbial);
Issuing authority (e.g. WHO, BCR, IRMM, USP, ATCC);
Records of characterization (if applicable); *
Molecular form(s) of, or surrogate for, the analyte (e.g. steric isomer
for an amino acid, or glycerol for glycerol ester);
Method used to establish value (AL);
Matrix (e.g. buffered bovine albumin solution, analyte concentration
in water, or soil, analyte concentration in air);
State(s) of aggregation (gas, liquid, solid);
Phase(s) (solution, suspension, lyophilized);
Records of use (required); *
A2LA
Document Issued:
Page 12 of 14

Comments
Y N NA
5.9 Assuring the Quality of Testing and Calibration Results

5.9.1 When commercial proficiency testing programs are available,
laboratories shall participate in these programs annually for all
targets, unless the programs are not relevant to the testing
performed.
Often, proficiency testing is not available for specific threat agents
tested. In the event an external proficiency testing program is not
available, the laboratory shall demonstrate competency annually,
using blind spiked samples (spiked with target or surrogate).
Prior to achieving initial accreditation, the laboratory shall have
successfully participated in one of the two requirements listed above
for each method on their desired Scope.
Table 1. Validation Recommendations for Chemistry Methods

Originating Laboratory Study – Level 1, 2, 3, 4 (Refer to R231
for details)
Appropriate # of matrices for selected level
Appropriate # of sources for selected level
Appropriate # participating laboratories for selected level
Appropriate # minimum analyte level for selected level
Replicates per matrix tested at each level
Table 2. Validation Recommendations for Microbiological
Methods
A2LA
Document Issued:
Page 13 of 14

Comments
Y N NA
Originating Laboratory Study – Level 1, 2, 3, 4 (Refer R231 for

details)
Appropriate # of strains of target organism (inclusivity)
Appropriate # of strains of non-target organism (exclusivity)
Appropriate # of matrices
Appropriate # of analyte levels/matrix
Appropriate # of replicates per matrix
Aging of inoculated samples prior to testing
Addition of competitor strain
Comparison to recognized method
Table 3. Validation Recommendations for Radiological Methods
Originating Laboratory Study – Level 1, 2, 3, 4(Refer to R231
for details)
Appropriate # of matrices
Appropriate # of sources for selected level
Appropriate # participating laboratories
Appropriate # minimum analyte levels/matrix
Appropriate # of replicates per matrix tested at each level
Document Revision History
Date Description
4/1/2016 Initial publication.
A2LA
Document Issued:
Page 14 of 14
8/23/17  Revised opening paragraph.

 Formatting throughout.
 4.13.2.1: added additional wording.
 5.4: Removed source of validation requirements.
 5.4.5.3.2: Added additional clause numbers to further distinguish clauses from each other.
 5.4.5.3.3: Removed reference to validation tools.
 5.4.5.3.5: Added additional clause numbers to further distinguish clauses from each other.
 5.4.6: Revised clause numbers.
 5.6.1: Added additional clause numbers to further distinguish clauses from each other.
 5.6.1.3: Removed repeated clause.
 5.6.3.2: Changed reference from G34 to ISO/IEC 17034.
 5.9.1: Split clause into three separate assessable clauses.

conversion/tmp/activity - Task - Scratch/582676692.docx: Document Issued: 12/6/17 Page 1 of 14

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

conversion/tmp/activity - Task - Scratch/582676692.docx: Document Issued: 12/6/17 Page 1 of 14

Uploaded by

Copyright:

Available Formats

A2LA

Master Code: Assessment ID:

{RESERVED FOR A2LA ASSESSORS ONLY}

Requirement Reference Compliance

4.0 Management Requirements

{RESERVED FOR A2LA ASSESSORS ONLY}

Requirement Reference Compliance

Identification of equipment used that is critical to the process (ex.

{RESERVED FOR A2LA ASSESSORS ONLY}

Requirement Reference Compliance

5.4.5.1 Prior to developing the proposed methods:

{RESERVED FOR A2LA ASSESSORS ONLY}

Requirement Reference Compliance

5.4.5.2.3 Modification of a method’s range beyond validated levels;

{RESERVED FOR A2LA ASSESSORS ONLY}

Requirement Reference Compliance

{RESERVED FOR A2LA ASSESSORS ONLY}

Requirement Reference Compliance

Statistics: Statistical techniques are employed to evaluate accuracy,

{RESERVED FOR A2LA ASSESSORS ONLY}

Requirement Reference Compliance

5.4.5.3.5.1 The following practices shall include, but not be limited

{RESERVED FOR A2LA ASSESSORS ONLY}

Requirement Reference Compliance

{RESERVED FOR A2LA ASSESSORS ONLY}

Requirement Reference Compliance

Molecular form(s) of or surrogate for the analyte (e.g. steric isomer

{RESERVED FOR A2LA ASSESSORS ONLY}

Requirement Reference Compliance

5.9 Assuring the Quality of Testing and Calibration Results

Table 1. Validation Recommendations for Chemistry Methods

{RESERVED FOR A2LA ASSESSORS ONLY}

Requirement Reference Compliance

Originating Laboratory Study – Level 1, 2, 3, 4 (Refer R231 for

Document Revision History

8/23/17  Revised opening paragraph.

You might also like