en
Anti-CCP
Anti-CCP Reagent Kit
Read Highlighted Changes: Revised March 2018.
Instructions must be carefully followed. Reliability of assay results
cannot be guaranteed if there are any deviations from these
instructions.
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NAME
Alinity i Anti-CCP Reagent Kit
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INTENDED USE
The Alinity i Anti-CCP assay is a chemiluminescent microparticle
immunoassay (CMIA) used for the semi-quantitative determination of
the IgG class of autoantibodies specific to cyclic citrullinated peptide
(CCP) in human serum or plasma on the Alinity i analyzer.
The Alinity i Anti-CCP assay is to be used as an aid in the diagnosis
of Rheumatoid Arthritis (RA) and should be used in conjunction
with other clinical information. Autoantibody levels represent one
parameter in a multicriterion diagnostic process, encompassing both
clinical and laboratory-based assessments.
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SUMMARY AND EXPLANATION OF THE TEST
Rheumatoid Arthritis (RA) is a common, systemic autoimmune
disease affecting 0.5-1% of the population. It is characterized by
chronic inflammation of the synovium, which commonly leads to
progressive joint destruction and in most cases, to disability and
reduction of quality of life.1 Evidence gained over the last few years
suggests that aggressive therapy given early in the disease has the
greatest therapeutic potential.2, 3
The serum of RA patients contains a variety of antibodies
directed against self-antigens. The most widely known of these
autoantibodies is the rheumatoid factor (RF) antibody directed
against the constant domain of IgG molecules. The presence
of RF is one of the American College of Rheumatology's (ACR)
criteria for the classification of RA.4 Although the RF test has good
sensitivity for RA, it is not very specific for the disease as it can
also be detected in the serum of patients with other rheumatic
or inflammatory diseases and even in a substantial percentage
of the healthy (elderly) population.5 For several years it has been
recognized that antibodies to anti-perinuclear factor (APF) and
anti-keratin (AKA) are highly specific for RA. It was subsequently
reported that both of these antibodies reacted with native filaggrin
and are now referred to as anti-filaggrin antibodies (AFA).6-8 More
recently it has been shown that all of these antibodies are directed
to citrulline-containing epitopes.9 Citrulline is a non-standard amino
acid, as it is not incorporated into proteins during protein synthesis.
It can, however, be generated via post-translational modification of
arginine residues by the enzyme peptidyl arginine deiminase (PAD).10
In 1998, Schellekens and colleagues reported that linear peptides
containing citrulline (CP) were very specific for RA antibodies (96%)
in an ELISA based assay.11 Subsequent work demonstrated that
cyclic variants of these peptides, termed cyclic citrullinated peptides
(CCP), were equally specific for RA, but with a higher sensitivity
than linear peptides.12 To improve the sensitivity of the CCP test
further, several dedicated libraries of citrulline-containing peptides
were screened with RA sera and a new set of peptides (CCP2)
were discovered which gave superior performance compared to the
CCP1 test.13 Over the last few years, many independent studies
have confirmed the diagnostic performance of the CCP2 test.14, 15 In
2007, the European League against Rheumatism (EULAR) published
guidelines for the diagnosis of early RA, and the measurement of
antibodies to anti-CCP was included as a serology marker.16
1
09P27
ABBL502R02
B9P270
09P2720
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BIOLOGICAL PRINCIPLES OF THE PROCEDURE
This assay is a two-step immunoassay with an automated sample
pretreatment for the semi-quantitative determination of the IgG
class of autoantibodies specific to cyclic citrullinated peptide (CCP)
in human serum or plasma using chemiluminescent microparticle
immunoassay (CMIA) technology.
Sample and wash buffer are combined. An aliquot of the prediluted
sample, CCP coated paramagnetic microparticles, and sample
diluent are combined and incubated. The anti-CCP antibodies
present in the sample bind to the CCP coated microparticles. The
mixture is washed. Anti-human IgG acridinium-labeled conjugate is
added to create a reaction mixture and incubated. Following a wash
cycle, Pre-Trigger and Trigger Solutions are added.
The resulting chemiluminescent reaction is measured as relative
light units (RLUs). There is a direct relationship between the amount
of anti-CCP antibody in the sample and the RLUs detected by the
system optics.
For additional information on system and assay technology, refer to
the Alinity ci-series Operations Manual, Section 3.
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REAGENTS
Kit Contents
Alinity i Anti-CCP Reagent Kit 09P27
Volumes (mL) listed in the table below indicate the volume per
cartridge.
09P2720
Tests per cartridge 100
Number of cartridges per kit 2
Tests per kit 200
6.6 mL
6.1 mL
10.4 mL
 CCP coated microparticles in phosphate buffer with
protein (bovine) stabilizer and surfactant. Minimum concentration:
0.05% solids. Preservative: sodium azide.
 Mouse anti-human IgG: acridinium-labeled conjugate in
MES buffer with protein (bovine) stabilizer and surfactant. Minimum
concentration: 10 ng/mL. Preservatives: Nipasept and Sarafloxacin.
 Phosphate buffer with protein (bovine) stabilizer and
surfactant. Preservative: sodium azide.
Warnings and Precautions
•
• For In Vitro Diagnostic Use
•
Safety Precautions
CAUTION: This product requires the handling of human specimens.
It is recommended that all human-sourced materials be considered
potentially infectious and handled in accordance with the OSHA
Standard on Bloodborne Pathogens. Biosafety Level 2 or other
appropriate biosafety practices should be used for materials that
contain or are suspected of containing infectious agents.17-20
 Indications of Reagent Deterioration
The following warnings and precautions apply to:
and Deterioration of the reagents may be indicated when a calibration
error occurs or a control value is out of the specified range.
Contains sodium azide.
Associated test results are invalid, and samples must be retested.
EUH032 Contact with acids liberates very toxic gas.
Assay recalibration may be necessary.
P501 Dispose of contents / container in
For troubleshooting information, refer to the Alinity ci-series
accordance with local regulations.
Operations Manual, Section 10.
Safety Data Sheets are available at www.abbottdiagnostics.com or
contact your local representative.
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INSTRUMENT PROCEDURE
The Alinity i Anti-CCP assay file must be installed on the Alinity i
For a detailed discussion of safety precautions during system analyzer prior to performing the assay.
operation, refer to the Alinity ci-series Operations Manual, Section 8.
For detailed information on assay file installation and viewing and
Reagent Handling editing assay parameters, refer to the Alinity ci-series Operations
• Upon receipt, gently invert the unopened reagent kit by rotating Manual, Section 2.
it over and back for a full 180 degrees, 5 times with green label For information on printing assay parameters, refer to the Alinity ci-
stripe facing up and then 5 times with green label stripe facing series Operations Manual, Section 5.
down. This ensures that liquid covers all sides of the bottles For a detailed description of system procedures, refer to the Alinity
within the cartridges. During reagent shipment, microparticles ci-series Operations Manual.
can settle on the reagent septum.
–– Place a check in the square on the reagent kit to indicate to ll
SPECIMEN COLLECTION AND PREPARATION
others that the inversions have been completed. FOR ANALYSIS
• After mixing, place reagent cartridges in an upright position for Specimen Types
1 hour before use to allow bubbles that may have formed to The specimen types listed below were verified for use with this
dissipate. assay.
• If a reagent cartridge is dropped, place in an upright position Other specimen types and collection tube types have not been
for 1 hour before use to allow bubbles that may have formed to verified with this assay.
dissipate. Specimen Types Collection Tubes
• Reagents are susceptible to the formation of foam and bubbles. Serum Serum
Bubbles may interfere with the detection of the reagent level in Serum separator
the cartridge and cause insufficient reagent aspiration that may
Plasma Lithium heparin plasma separator
adversely affect results.
Potassium EDTA
For a detailed discussion of reagent handling precautions during
system operation, refer to the Alinity ci-series Operations Manual, • Performance has not been established for the use of cadaveric
Section 7. specimens or the use of body fluids other than human serum or
Reagent Storage plasma.
• Liquid anticoagulants may have a dilution effect resulting in lower
Storage Maximum Additional Storage
Temperature Storage Time Instructions concentration values for individual specimens.
Unopened 2 to 8°C Until Store in upright position. • Plasma specimens from different anticoagulant tube types
expiration If cartridge does not should not be used interchangeably for monitoring anti-CCP.
date remain upright, gently • The instrument does not provide the capability to verify specimen
invert the cartridge 10 types. It is the responsibility of the operator to verify that the
times and place in an correct specimen types are used in the assay.
upright position for 1 hour Specimen Conditions
before use. • Do not use:
Onboard System 30 days –– heat-inactivated specimens
Temperature –– pooled specimens
Opened 2 to 8°C Until Store in upright position. –– grossly hemolyzed specimens
expiration If cartridge does not –– specimens with obvious microbial contamination
date remain upright during
• For accurate results, serum and plasma specimens should be
storage, discard the
free of fibrin, red blood cells, and other particulate matter. Serum
cartridge.
specimens from patients receiving anticoagulant or thrombolytic
Do not reuse original therapy may contain fibrin due to incomplete clot formation.
reagent caps or
• To prevent cross contamination, use of disposable pipettes or
replacement caps due to
pipette tips is recommended.
the risk of contamination
and the potential to Preparation for Analysis
compromise reagent • Follow the tube manufacturer’s processing instructions for
performance. collection tubes. Gravity separation is not sufficient for specimen
preparation.
Reagents may be stored on or off the system. If removed from the
• Specimens should be free of bubbles. Remove bubbles with an
system, store reagents with new replacement caps in an upright
applicator stick before analysis. Use a new applicator stick for
position at 2 to 8°C. For reagents stored off the system, it is
each specimen to prevent cross-contamination.
recommended that they be stored in their original trays or boxes to
To ensure consistency in results, recentrifuge specimens prior to
ensure they remain upright.
testing if
For information on unloading reagents, refer to the Alinity ci-series
• they contain fibrin, red blood cells, or other particulate matter
Operations Manual, Section 5.
• they require repeat testing.

2
NOTE: If fibrin, red blood cells, or other particulate matter are If testing will be delayed more than 22 hours for specimens stored
observed, mix by low speed vortex or by inverting 10 times prior to at room temperature or more than 7 days for specimens stored at
recentrifugation. 2 to 8°C, remove serum or plasma from the clot, red blood cells, or
Prepare frozen specimens as follows: separator gel and store at -20°C or colder.
• Frozen specimens must be completely thawed before mixing. Avoid more than three freeze/thaw cycles.
• Mix thawed specimens thoroughly by low speed vortex or by Specimen Shipping
inverting 10 times. Package and label specimens in compliance with applicable state,
• Visually inspect the specimens. If layering or stratification is federal, and international regulations covering the transport of clinical
observed, mix until specimens are visibly homogeneous. specimens and infectious substances.
• If specimens are not mixed thoroughly, inconsistent results may
be obtained.
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PROCEDURE
• Recentrifuge specimens. Materials Provided
Recentrifugation of Specimens 09P27 Alinity i Anti-CCP Reagent Kit
• Transfer specimens to a centrifuge tube and centrifuge at Materials Required but not Provided
100 000 g-minutes. • Alinity i Anti-CCP assay file
• Examples of acceptable time and force ranges that meet this • 09P2701 Alinity i Anti-CCP Calibrators
criterion are listed in the table below. • 09P2710 Alinity i Anti-CCP Controls or other control material
Centrifugation time using alternate RCF values can be calculated • Alinity Trigger Solution
using the following formula: • Alinity Pre-Trigger Solution
100 000 g-minutes • Alinity i-series Concentrated Wash Buffer
Minimum Centrifugation time (minutes) =
RCF For information on materials required for operation of the instrument,
Recentrifugation Time refer to the Alinity ci-series Operations Manual, Section 1.
(Minutes) RCF (x g)* g-Minutes For information on materials required for maintenance procedures,
10 10 000 100 000 refer to the Alinity ci-series Operations Manual, Section 9.
20 5000 100 000
Assay Procedure
40 2500 100 000
For a detailed description of how to run an assay, refer to the Alinity
* To ensure consistency in results, specimens must be centrifuged ci-series Operations Manual, Section 5.
using an appropriate tube at a minimum of 2500 RCF to obtain a • If using primary or aliquot tubes, refer to the Alinity ci-series
minimum of 100 000 g-minutes. Operations Manual, Section 4 to ensure sufficient specimen is
RCF = 1.12 × rmax (rpm/1000)2 present.
RCF - The relative centrifugal force generated during • To minimize the effects of evaporation, verify adequate sample
centrifugation. cup volume is present prior to running the test.
rpm - The revolutions per minute of the rotor on which • Maximum number of replicates sampled from the same sample
the specimens are being spun (usually the digital cup: 10
readout on the centrifuge will indicate the rpm). –– Priority:
Centrifugation The time should be measured from the time the ◦◦ Sample volume for first test: 60 µL
Time - rotor reaches the required RCF or rpm to the time it ◦◦ Sample volume for each additional test from same sample
begins decelerating. cup: 10 µL
rmax - Radius of the rotor in millimeters. NOTE: If custom –– ≤ 3 hours on the reagent and sample manager:
tube adapters (i.e., adapters not defined by the
◦◦ Sample volume for first test: 150 µL
centrifuge manufacturer) are used, then the radius
(rmax) should be manually measured in millimeters ◦◦ Sample volume for each additional test from same sample
and the RCF calculated. cup: 10 µL
g-minutes - The unit of measure for the product of RCF (× g) –– > 3 hours on the reagent and sample manager:
and centrifugation time (minutes). ◦◦ Replace with a fresh aliquot of sample.
• Refer to the Alinity i Anti-CCP calibrator package insert and/
• Transfer clarified specimen to a sample cup or secondary tube or Alinity i Anti-CCP control package insert for preparation and
for testing. For centrifuged specimens with a lipid layer, transfer usage.
only the clarified specimen and not the lipemic material.
• For general operating procedures, refer to the Alinity ci-series
Specimen Storage Operations Manual, Section 5.
Maximum • For optimal performance, it is important to perform routine
Specimen Storage maintenance as described in the Alinity ci-series Operations
Type Temperature Time Special Instructions
Manual, Section 9. Perform maintenance more frequently when
Serum/ Room 22 hours Specimens may be required by laboratory procedures.
Plasma temperature stored on or off the
(study clot, red blood cells, or Sample Dilution Procedures
performed at separator gel. Samples with anti-CCP value exceeding 195.6 U/mL are flagged
30°C) with the code "> 195.6 U/mL" and may be diluted using either the
Automated Dilution Protocol or the Manual Dilution Procedure.
2 to 8°C 7 days Specimens may be
Automated Dilution Protocol
stored on or off the
clot, red blood cells, or The system performs a 1:6 dilution of the sample and automatically
separator gel. calculates the concentration by multiplying the result by the dilution
factor.

3
Manual Dilution Procedure
Suggested dilution: 1:10
Add 50 μL of the sample to 450 μL of Alinity i Anti-CCP Negative
Control.
Avoid contamination of Negative Control when transferring volume for
manual dilution.
Refer to the Alinity i Anti-CCP control package insert for preparation
and storage.
The operator must enter the dilution factor in the Specimen or
Control tab of the Create Order screen. The system will use this
dilution factor to automatically calculate the concentration of the
sample and report the result.
If the operator does not enter the dilution factor, the result must be
manually multiplied by the appropriate dilution factor before reporting
the result. If a diluted sample result is less than the lower value of
the measuring interval of 0.5 U/mL, do not report the result. Rerun
using an appropriate dilution.
For detailed information on ordering dilutions, refer to the Alinity ci-
series Operations Manual, Section 5.
Calibration
For instructions on performing a calibration, refer to the Alinity ci-
series Operations Manual, Section 5.
Each assay control must be tested to evaluate the assay calibration.
Once a calibration is accepted and stored, all subsequent samples
may be tested without further calibration unless:
• A reagent kit with a new lot number is used.
• Daily quality control results are outside of statistically-based
quality control limits used to monitor and control system
performance, as described in the Quality Control Procedures
section of this package insert.
–– If statistically-based quality control limits are not available,
then the calibration should not exceed a 30-day limit for
recalibration frequency.
This assay may require recalibration after maintenance to critical
parts or subsystems or after service procedures have been
performed.
Quality Control Procedures
The recommended control requirement for the Alinity i Anti-CCP
assay is that a single sample of each control level be tested once
every 24 hours each day of use.
Additional controls may be tested in accordance with local, state,
and/or federal regulations or accreditation requirements and your
laboratory’s quality control policy.
To establish statistically-based control limits, each laboratory should
establish its own concentration target and ranges for new control lots
at each clinically relevant control level. This can be accomplished
by assaying a minimum of 20 replicates over several (3-5) days and
using the reported results to establish the expected average (target)
and variability about this average (range) for the laboratory. Sources
of variation that should be included in this study in order to be
representative of future system performance include:
• Multiple stored calibrations
• Multiple reagent lots
• Multiple calibrator lots
• Multiple processing modules (if applicable)
• Data points collected at different times of the day
Refer to published guidelines for information or general control
recommendation, for example Clinical and Laboratory Standards
Institute (CLSI) Document C24-A3 or other published guidelines, for
general quality control recommendations.21
• If more frequent control monitoring is required, follow the
established quality control procedures for your laboratory.
4
• If quality control results do not meet the acceptance criteria
defined by your laboratory, sample results may be suspect.
Follow the established quality control procedures for your
laboratory. Recalibration may be necessary. For troubleshooting
information, refer to the Alinity ci-series Operations Manual,
Section 10.
• Review quality control results and acceptance criteria following a
change of reagent or calibrator lot.
Quality Control Guidance
Refer to “Basic QC Practices” by James O Westgard, Ph.D. for
guidance on laboratory quality control practices.22
Verification of Assay Claims
For protocols to verify package insert claims, refer to Verification of
Assay Claims in the Alinity ci-series Operations Manual.
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RESULTS
Calculation
The Alinity i Anti-CCP assay utilizes a 4 Parameter Logistic Curve fit
data reduction method (4PLC, Y-weighted) to generate a calibration
and results.
Flags
Some results may contain information in the Flags field. For a
description of the flags that may appear in this field, refer to the
Alinity ci-series Operations Manual, Section 5.
Measuring Interval
Measuring interval is defined as the range of values in U/mL which
meets the limits of acceptable performance for linearity, imprecision,
and bias.
The measuring interval of the Alinity i Anti-CCP assay is 0.5 to
195.6 U/mL.
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LIMITATIONS OF THE PROCEDURE
• For diagnostic purposes, the Alinity i Anti-CCP results should
be used in conjunction with other clinical data; e.g., symptoms,
medical history, etc.
• If the Alinity i Anti-CCP results are inconsistent with clinical
evidence, additional testing is suggested to confirm the result.
• The value of anti-CCP in juvenile arthritis has not been
determined.
• Some specimens may not dilute linearly because of
the heterogeneity of the autoantibodies with respect to
physiochemical properties.
• Alinity i Anti-CCP results should not be used interchangeably
with other manufacturers' methods for anti-CCP
determinations.
• Plasma specimens from different anticoagulant tube types
should not be used interchangeably for monitoring anti-CCP.
• Specimens from patients who have received preparations of
mouse monoclonal antibodies for diagnosis or therapy may
contain human anti-mouse antibodies (HAMA).23, 24 Specimens
containing HAMA may produce anomalous values when tested
with assay kits such as Alinity i Anti-CCP that employ mouse
monoclonal antibodies.23
• Heterophilic antibodies in human specimens can react
with reagent immunoglobulins, interfering with in vitro
immunoassays.25 Patients routinely exposed to animals or to
animal serum products can be prone to this interference and
anomalous values may be observed. Additional information may
be required for diagnosis.
• Refer to the SPECIMEN COLLECTION AND PREPARATION FOR
ANALYSIS section of this package insert for specimen limitations.
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EXPECTED VALUES Analytical Specificity
This study was performed on the ARCHITECT i System. This study was performed on the ARCHITECT i System.
Representative performance data are provided in this section. To assess the potential cross-reactivity of the CCP antigen used
Results obtained in individual laboratories may vary. in the ARCHITECT Anti-CCP assay with other autoantibodies, the
It is recommended that each laboratory determine its own reference assay was evaluated with 20 samples positive for various other
range based upon its particular locale and population characteristics. autoantibodies and negative for CCP antibodies. The following
In a representative study, serum specimens from 199 asymptomatic, autoantibodies (1-4 samples of each) were tested in the assay: SSA,
apparently healthy males (n = 126) and females (n = 73), with SSB, Sm, RNP, ds-DNA, Jo-1, Scl-70, Ribo-P, TPO, ANA, and AMA.
an age range of 19 to 67 years, were tested with the ARCHITECT The study showed no significant cross-reactivity of the CCP antigen
Anti-CCP assay. No differences attributable to gender or age were with any of these other autoantibodies.
observed. Specimen values ranged from < 0.5 U/mL to 2.5 U/mL. Interference
A cut-off of 5.0 U/mL was chosen, whereby a result of ≥ 5.0 U/mL This study was performed on the ARCHITECT i System.
is considered positive and a result of < 5.0 U/mL is considered Potentially Interfering Endogenous Substances
negative. A study was performed based on guidance from CLSI EP07-A2.29
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SPECIFIC PERFORMANCE CHARACTERISTICS Serum samples with anti-CCP levels across the assay range of
Representative performance data are provided in this section. 0.5 U/mL to 200.0 U/mL were supplemented with the potentially
Results obtained in individual laboratories may vary. interfering compounds listed in the table below. The maximum
The Alinity i analyzer and the ARCHITECT i System utilize the same deviation of anti-CCP concentration observed in serum samples
reagents and sample/reagent ratios. during these studies ranged from:
Unless otherwise specified, all studies were performed on the Alinity • -7.6% to 0.8% for anti-CCP concentrations > 10.0 U/mL
i analyzer. • -1.0% to 7.5% for anti-CCP concentrations ≥ 5.0 U/mL to
≤ 10.0 U/mL
Precision
• -0.3 U/mL to 0.2 U/mL for anti-CCP concentrations < 5.0 U/mL
Within-Laboratory Precision
Potentially Interfering Substance Interferent Level
A study was performed based on guidance from CLSI EP05-A2.26
Bilirubin 20 mg/dL
Testing was conducted using 1 lot of the Alinity i Anti-CCP Reagent
Hemoglobin 800 mg/dL
Kit, 1 lot of the Alinity i Anti-CCP Calibrators, and 1 lot of the Alinity
i Anti-CCP Controls and 1 instrument. One control and 6 human Total Protein 12 g/dL
serum panels were assayed in a minimum of 2 replicates at 2 Triglycerides 3000 mg/dL
separate times per day on 20 different days. Rheumatoid Factor 200 IU/mL
Within-Run Within-Laboratory Tube Type Matrix Comparison
(Repeatability) (Total)a
Sample n Mean (U/mL) SD %CV SD %CV
This study was performed on the ARCHITECT i System.
Positive Control 80 21.5 0.41 1.9 0.45 2.1 The specimen collection tubes listed below were verified for use with
Panel 1 80 10.1 0.16 1.5 0.18 1.8 the ARCHITECT Anti-CCP assay.
Panel 2 80 30.2 0.40 1.3 0.70 2.3 • serum, serum separator, lithium heparin plasma separator, and
Panel 3 80 66.7 1.18 1.8 1.56 2.3
potassium EDTA.
Panel 4 80 138.6 3.90 2.8 5.67 4.1 When compared to the control tube type (serum), the tube types
Panel 5 80 3.0 0.06 1.9 0.06 2.1
evaluated for samples with anti-CCP values < 5.0 U/mL showed less
than a 0.5 U/mL difference on average, and the tube types evaluated
Panel 6 80 183.4 8.83 4.8 9.84 5.4
for samples with anti-CCP values ranging from 5.3 to 178.8 U/mL
a Includes within-run, between-run, and between-day variability. showed less than a 10% difference on average. The distribution of
Lower Limits of Measurement the differences or percent differences per tube type is listed in the
following table.
A study was performed based on guidance from CLSI EP17-A2.27
Testing was conducted using 3 lots of the Alinity i Anti-CCP Reagent Distribution of Absolute Distribution of Absolute Percent
Differences < 0.5 U/mL Differences for Samples with Anti-CCP
Kit on each of 2 instruments over a minimum of 3 days. The Values 5.3 to 178.8 U/mL
for Samples with Anti-
maximum observed Limit of Blank (LoB), Limit of Detection (LoD), Tube Type CCP Values < 5.0 U/mL < 10% ≥ 10% to ≤ 20% > 20%
and Limit of Quantitation (LoQ) values are summarized below.
Serum Separator 100% 76% 16% 8%
U/mL (19/19) (19/25) (4/25) (2/25)
LoBa 0.1 Potassium EDTA 100% 72% 24% 4%
LoDb 0.3 (19/19) (18/25) (6/25) (1/25)
LoQc 0.4 Lithium Heparin 100% 80% 16% 4%
a Plasma (19/19) (20/25) (4/25) (1/25)
The LoB represents the 95th percentile from n ≥ 60 replicates of Separator
zero-analyte samples.
b The LoD represents the lowest concentration at which the analyte
Method Comparison
can be detected with 95% probability based on n ≥ 60 replicates of A study was performed based on guidance from CLSI EP09-A330
low-analyte level samples. using the Passing-Bablok regression method.
c The LoQ was determined from n ≥ 60 replicates of low-analyte
Correlation Concentration
level samples and is defined as the lowest concentration at which a Units n Coefficient Intercept Slope Range
maximum allowable precision of 20 %CV was met. Alinity i Serum U/mL 120 0.99 0.08 0.94 0.5-183.1
Linearity Anti-CCP vs
ARCHITECT
A study was performed based on guidance from CLSI EP06-A.28 Anti-CCP
This assay is linear across the measuring interval of 0.5 to
195.6 U/mL.

5
Negative and Positive Percent Agreement
Cut-off
Using a cut-off of 5.0 U/mL for the ARCHITECT Anti-CCP assay, the
concordance was calculated to be 99.3%.
Percent Agreement
A study was performed based on guidance from CLSI EP12-A2.31
ARCHITECT Anti-CCP
Positive Negative
Alinity i Anti-CCP Positive 119
Negative 0 108
Positive % agreement = 100.00% (119/119); 95% confidence interval
(96.95 - 100.00%)
Negative % agreement = 100.00% (108/108); 95% confidence interval
(96.64 - 100.00%)
Receiver Operator Characteristic (ROC) Analysis
A ROC analysis was carried out using ARCHITECT Anti-CCP data.
The area under the curve (AUC) for the ARCHITECT Anti-CCP assay
was 0.873 (95% confidence interval: 0.849-0.897). The ROC analysis
curve is shown below.
x 1 - Specificity (false positives) No discrimination
y Sensitivity (true positives) ARCHITECT
High Dose Hook
This study was performed on the ARCHITECT i System.
High dose hook is a phenomenon whereby very high level specimens
may read within the dynamic range of the assay. No high dose
hook effect was observed when a sample containing approximately
2000 U/mL of anti-CCP antibody was assayed.
Clinical Sensitivity and Specificity
This study was performed on the ARCHITECT i System.
The clinical sensitivity was determined for 496 confirmed RA
individuals, and clinical specificity was determined for 499 non-
RA specimens (299 from patients with other rheumatic and
non-rheumatic disorders and 200 from asymptomatic apparently
healthy individuals). Using a cut-off of 5.0 U/mL, the sensitivity was
calculated to be 70.6% with a specificity of 98.2%. The results are
summarized in the following tables.
ARCHITECT Anti-CCP
Specimen Category Total n Positive n % Sensitivity
Confirmed RAa 496 350 70.6
a RA patients were classified according to the ACR Criteria.4
ARCHITECT Anti-CCP
Specimen Category Total n Positive n % Specificity
Non-RA Specimens 499 9 98.2
in Total
Non-RA Healthy 200 1 99.5
Asymptomatic
Non-RA Disease 299 8 97.3
Specimensa
aThe non-RA diseases were Ankylosing Spondylitis, Autoimmune
0
Thyroiditis/Hashimoto's Disease, Crohn's Disease, Dermatomyositis,
Epstein-Barr Virus, Lyme Disease, Osteoarthritis, Polymyalgia
Rheumatica, Polymyositis, Psoriatic Arthritis, Reactive Arthritis/
Reiter's Syndrome, Scleroderma, Sjögren's Syndrome, Systemic
Lupus Erythematosus, and Ulcerative Colitis.
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BIBLIOGRAPHY
1. Feldmann M, Brennan FM, Maini RN. Rheumatoid arthritis. Cell
1996;85:307-310
2. Landewé RB. The benefits of early treatment in rheumatoid arthritis:
confounding by indication, and the issue of timing. Arthritis Rheum
2003;48(1):1-5.
3. Lard LR, Visser H, Speyer I, et al. Early versus delayed treatment
in patients with recent-onset rheumatoid arthritis: comparison of
two cohorts who received different treatment strategies. Am J Med
2001;111:446-451.
4. Arnett FC, Edworthy SM, Bloch DA, et al. The American Rheumatism
Association 1987 revised criteria for the classification of rheumatoid
arthritis. Arthritis Rheum 1988;31(3):315-324.
5. van Venrooij WJ, Hazes JM, Visser H. Anticitrullinated protein/
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Note for number formatting: For use by or on the order of a
• A space is used as thousands separator (example: 10 000 physician only (applicable to USA
specimens). classification only).
• A period is used to separate the integer part from the fractional Sample Diluent
part of a number written in decimal form (example: 3.12%).
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