The IMMULITE 2000 Anti-CCP IgG assay is an in vitro diagnostic immunoassay for the semi-quantitative determination of the IgG class of autoantibodies specific to cyclic citrullinated peptide (CCP) in human serum (including Serum Separator Tubes) or plasma (EDTA or lithium heparin) on the IMMULITE 2000 system. Detection of anti-CCP antibodies is 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 multi-criteria diagnostic process, encompassing both clinical and laboratory-based assessments.

The IMMULITE 2000 Anti-CCP IgG Calibration Verification Material (CVM) is for in vitro diagnostic use, as a control for calibration verification of the IMMULITE 2000 Anti-CCP IgG assay on the IMMULITE 2000 system.

Device Description

The IMMULITE 2000 Anti-CCP IgG assay consists of the following components:

Anti-CCP IgG bead pack coated with cyclic citrullinated peptide . (CCP) antigen
Anti-CCP IgG reagent wedge containing bovine calf intestine . conjugated to a monoclonal murine anti-human IgG antibody
Anti-CCP IgG adjustors, low and high, containing lyophilized . human serum with IgG reactive to CCP
Anti-CCP IgG controls, negative and positive, containing human . serum
. Autoantibody sample diluent containing protein/buffer matrix

The IMMULITE Anti-CCP IgG Calibration Verification Material consists of one set of four vials, containing low, intermediate and high levels of lyophilized human serum with IgG reactive to cyclic citrullinated peptide (CCP), in buffer with preservative, plus an anti-CCP-free sample.

AI/ML Overview

The IMMULITE® 2000 Anti-CCP IgG Assay is an in vitro diagnostic immunoassay for the semi-quantitative determination of the IgG class of autoantibodies specific to cyclic citrullinated peptide (CCP). It is used as an aid in the diagnosis of Rheumatoid Arthritis (RA) and should be used in conjunction with other clinical information.

Here's an analysis of the acceptance criteria and the study that proves the device meets those criteria:

1. Table of Acceptance Criteria and Reported Device Performance

The provided document does not explicitly state acceptance criteria in a single, consolidated table. However, performance characteristics are reported, which implicitly define what the manufacturer considers acceptable for their device to perform as intended and to demonstrate substantial equivalence to the predicate device. Based on the performance characteristics presented, the following can be inferred as the "acceptance criteria" through the demonstrated results.

Performance Characteristic	Acceptance Criteria (Implicit from Results)	Reported Device Performance
Precision (Total CV%)	Generally, Coefficients of Variation (CV%) for diagnostic assays should be within acceptable limits (e.g., <10-15% for clinically relevant concentrations).	Ranged from 4.3% to 14.8% for SERP2 (lowest conc.) across different samples and statistical breakdowns (Within-Run to Total Within-Lot). This is generally acceptable for immunoassay precision.
	Specifically, lower concentrations tend to have higher CVs.	Examples:
		- LPIC2 (47.4 U/mL): Total CV 4.3%
		- SERP2 (2.1-1.94 U/mL): Total CV 13.6% (20-Day) / 14.8% (Reproducibility)
		- SERP6 (144.0-139.18 U/mL): Total CV 4.8% (20-Day) / 5.1% (Reproducibility)
Linearity/Reportable Range	Demonstrating linearity across the specified reportable range (1.50 - 200 U/mL) with acceptable recovery.	Percent recovery generally within ±10% range (e.g., 77.4% to 104.3%). Most samples are above 90%. Exception: P13 (2.64 U/mL) at 77.4% recovery.
Detection Limit (LoD)	LoD should be sufficiently low for clinical utility.	LoD determined to be 1.46 U/mL. LoB 0.26 U/mL.
Analytical Specificity (Interference)	Mean interference of common endogenous substances should be less than a certain threshold (e.g., ±10%).	Mean interference for all tested substances (Albumin, Triglycerides, Hemoglobin, Bilirubin, Rheumatoid Factor) was less than 10%.
Method Comparison (vs. Predicate)	High positive and negative agreement with the predicate device.	Reagent Lot 1: Positive Agreement 86.9%; Negative Agreement 46.2%; Total Agreement 82.2%
		Reagent Lot 2: Positive Agreement 86.1%; Negative Agreement 42.3%; Total Agreement 81.6%
Clinical Sensitivity & Specificity	Demonstrating acceptable diagnostic performance (sensitivity and specificity) for aid in RA diagnosis.	Sensitivity: 63.6%; Specificity: 97.0%
Matrix Comparison	High correlation and minimal bias between different sample types (serum, plasma).	Correlation Coefficient: 1.000 for all comparisons (Lithium Heparin, SST Serum, EDTA Plasma vs. Serum). Slopes near 1, intercepts near 0.

2. Sample Sizes Used for the Test Set and Data Provenance

The document describes several studies, each with its own sample size and data provenance:

Precision/Reproducibility Study:
- 20-Day Imprecision: 11 samples over 20 days, 2 instruments, 2 reagent lots, 2 runs/day, 2 replicates/sample. Total N for calculations is 320 for each sample. Data provenance not specified, but likely internal (manufacturer's lab).
- Reproducibility (Multi-site): Serum donor panels from precision study, 2 different reagent lots, run over 10 days, 2 runs/day, 4 replicates/run. Tested at three external testing sites. N=244 to 246 for each sample for Reagent Lot 201, and N=228 to 240 for Reagent Lot 202. Data provenance suggests multi-site, potentially from different geographical regions but not explicitly stated. Retrospective.
Linearity Study: A dilution series (11 dilutions) prepared from a reactive and non-reactive serum pool. Run in triplicate on one IMMULITE® 2000 instrument. Data provenance likely internal.
Detection Limit Study:
- LoB: Non-reactive donor sample tested on 3 instruments over 5 days, twice daily, 2 lots of reagent, 2 replicates/run. NB = 732 blank measurements. Data provenance not specified, likely internal.
- LoD: Five Anti-CCP serum samples (0.26 to 2.5 U/mL) assayed in replicates of 6 using 2 reagent kit lots, run for 5 days with 2 runs/day. Two instruments and 2 operators, generating 960 observations. Data provenance not specified, likely internal.
Analytical Specificity (Interference) Study: One control sample and five (5) different sample pools with differing Anti-CCP concentrations (range 2.0 U/mL to 205 U/mL) were used. Each substance spiked separately. Data provenance not specified, likely internal.
Assay Cut-off Determination: 388 samples: 212 apparently healthy samples, 106 RA positive samples, 70 anti-CCP positive samples (as determined by a 3rd party method). Data provenance not specified (country of origin), but implies external patient samples. Retrospective.
Method Comparison with Predicate Device:
- Reagent Lot 1: 255 unaltered patient serum samples.
- Reagent Lot 2: 256 unaltered patient serum samples.
- Data provenance not specified (country of origin), but implies external patient samples. Retrospective.
Matrix Comparison: 39 matched serum and plasma samples (serum clot, Lithium Heparin, SST, EDTA plasma tubes). 21 of these were spiked. Data provenance not specified, likely internal or from a single collection site.
Clinical Sensitivity and Specificity Study: 1512 patient serum samples. 1048 RA positive (classified by ACR criteria) and 464 non-RA (with potentially cross-reactive diseases). Data provenance not specified (country of origin), but implies external patient samples. Retrospective.
Expected Values/Reference Range Study: 200 serum samples from presumed healthy male and female donors. Data provenance not specified (country of origin). Retrospective.

3. Number of Experts Used to Establish Ground Truth for the Test Set and Their Qualifications

The document does not specify the number of experts used or their qualifications for establishing the ground truth for any of the test sets.

For the Assay Cut-off Determination and Method Comparison studies, "RA positive samples" and "anti-CCP positive samples, as determined by a 3rd party method" are mentioned, suggesting a clinical diagnosis or another validated assay served as the reference, but reviewer expertise is not detailed.
For the Clinical Sensitivity and Specificity Study, "RA positive patients were classified according to the ACR criteria." The American College of Rheumatology (ACR) criteria are a widely accepted standard for RA diagnosis, implying that these classifications were made by qualified clinicians (rheumatologists or physicians trained in RA diagnosis). However, the number of such experts and their specific qualifications are not provided.

4. Adjudication Method for the Test Set

The document does not describe any specific adjudication method (e.g., 2+1, 3+1, none) for the test sets.

For the Clinical Sensitivity and Specificity Study, the RA positive patient classification "according to the ACR criteria" is a standardized diagnostic approach, which implicitly involves clinical assessment. However, how consistency or discrepancies in these classifications were handled (e.g., by multiple clinicians, consensus panels) is not described.

5. If a Multi Reader Multi Case (MRMC) Comparative Effectiveness Study was Done

No, an MRMC comparative effectiveness study was not done. This device is an in vitro diagnostic (IVD) immunoassay, which generates quantitative or semi-quantitative results rather than images requiring human interpretation. Therefore, the concept of "human readers improving with AI vs. without AI assistance" is not applicable in this context. The comparison is primarily between the device's output and clinical reference standards or predicate IVD devices.

6. If a Standalone Study (Algorithm Only Without Human-in-the-Loop Performance) Was Done

Yes, the studies conducted are inherently "standalone" in the sense that they evaluate the performance of the IMMULITE® 2000 Anti-CCP IgG Assay directly, without a human-in-the-loop component for result generation. The assay produces a numerical result (U/mL) which is then interpreted against a clinical cut-off. Performance characteristics like precision, linearity, detection limit, analytical specificity, and clinical sensitivity/specificity assess the algorithm's (assay's) performance. While the interpretation of the results by clinicians for actual patient diagnosis involves human judgment, the device itself provides an objective measurement.

7. The Type of Ground Truth Used

Different types of ground truth were used depending on the study:

Clinical Sensitivity and Specificity: Patients classified as RA positive "according to the ACR criteria" (American College of Rheumatology criteria). This represents expert consensus clinical diagnosis based on established medical guidelines.
Assay Cut-off Determination: "RA positive samples" and "anti-CCP positive samples, as determined by a 3rd party method." This implies either clinical diagnosis (for RA positive) or results from a legally marketed predicate/reference assay (for anti-CCP positive).
Method Comparison with Predicate Device: The predicate device, DIASTAT™ Anti-Cyclic Citrullinated Peptide (anti-CCP) ELISA, results were used as the comparator for agreement analyses. This represents comparison to an existing, validated diagnostic method.
Laboratory Performance (Precision, Linearity, LoD, Interference): Ground truth is typically established by the inherent properties of the prepared samples (e.g., spiked concentrations, known non-reactive samples) or statistical calculations from repeated measurements.

8. The Sample Size for the Training Set

The document does not explicitly mention a "training set" in the context of machine learning or AI algorithms as the device is an immunoassay. However, if "training set" refers to samples used for developing and optimizing the assay's parameters (e.g., cut-off determination), the primary studies mentioned include:

Assay Cut-off determination: 388 samples were used (212 healthy, 106 RA positive, 70 anti-CCP positive from a 3rd party method) for ROC analysis to establish the cut-off. This set played a role analogous to a "training/optimization set" for the cut-off value.
Adjustors and Controls: "prepared in house and arbitrary units are assigned during the development process," suggesting internal work to define these.
Reference Calibrators: "A lot of reference calibrators was prepared by diluting the positive unit with negative normal human serum." These calibrators were "value-assigned by the dilution factor," and this internal standard is used for traceability.

9. How the Ground Truth for the Training Set Was Established

As noted above, for an immunoassay, the concept of a "training set" and its "ground truth" differs from typical AI/ML contexts.

Assay Cut-off Determination: The ground truth for the 388 samples used in ROC analysis for cut-off determination was established by:
- Clinical diagnosis: for "RA positive samples" and "healthy samples."
- Results from a 3rd party method: for "anti-CCP positive samples."
Reference Calibrators and Controls: These are established through internal arbitrary assignment, dilution factors, and extensive validation (e.g., precision, stability, value assignment across multiple lots and instruments) against an internal master batch (internal standard). This ensures consistency and reproducibility of the assay's quantitative output.

Summary

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IMMULITE® 2000 Anti-CCP IgG Assay

APR 0 4 2013

510(k) Summary as Required by 21 CFR 807.92

A. 510(k) Number	K121576
B. Purpose for Submission	New device
C. Measurand	Anti-cyclic citrullinated peptide (CCP) antibodies
D. Type of test	Semi-quantitative chemiluminescent immunometric assay
E. Applicant:	Siemens Healthcare Diagnostics Inc.511 Benedict AvenueTarrytown, NY 10591
F. Proprietary and EstablishedNames:	IMMULITE® 2000 Anti-CCP IgG Assay
G. RegulatoryInformation:

Regulation 21 CFR 866.5775 1

section:

Classification: Class II

1. Products Codes: NHX - Antibodies, Anti-Cyclic Citrullinated Peptide (CCP) JIT- Calibrators JJX – Single Analyte Control

Panel: Immunology (82)

The IMMULITE 2000 Anti-CCP IgG assay is an in vitro diagnostic H. Intended Use: immunoassay for the semi-quantitative determination of the IgG class of autoantibodies specific to cyclic citrullinated peptide (CCP) in human serum (including Serum Separator Tubes) or plasma (EDTA or lithium heparin) on the IMMULITE® 2000 system. Detection of anti-CCP antibodies is 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 multicriteria diagnostic process, encompassing both clinical and laboratorybased assessments.
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I. Device Description: The IMMULITE 2000 Anti-CCP IgG assay consists of the following components:

Anti-CCP IgG bead pack coated with cyclic citrullinated peptide . (CCP) antigen
Anti-CCP IgG reagent wedge containing bovine calf intestine . conjugated to a monoclonal murine anti-human IgG antibody
Anti-CCP IgG adjustors, low and high, containing lyophilized . human serum with IgG reactive to CCP
Anti-CCP IgG controls, negative and positive, containing human . serum
. Autoantibody sample diluent containing protein/buffer matrix

Predicate device name: DIASTATTM Anti-Cyclic Citrullinated Peptide (anti-CCP) ELISA

510(k) number: K023285

Comparison with predicate:

A comparison of the device features, intended use, laboratory data and other information demonstrates that the IMMULITE ® 2000 Anti-CCP IgG assay is substantially equivalent to the predicate device, DIASTATTM Anti-Cyclic Citrullinated Peptide (anti-CCP) ELISA, as summarized in the following tables.

J. Substantial Equivalence Information:

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	SIMILARITIES
Item	Device	Predicate
	IMMULITE 2000 Anti-CCP IgGAssay	DIASTATT™ Anti-CyclicCitrullinated Peptide (anti-CCP)ELISA
Intended Use	The IMMULITE® 2000 Anti-CCPIgG assay is an in vitro diagnosticimmunoassay for the semi-quantitative determination of theIgG class of autoantibodiesspecific to cyclic citrullinatedpeptide (CCP) in human serum orplasma on the IMMULITE®2000system. Detection of anti-CCPantibodies is used as an aid in thediagnosis of Rheumatoid Arthritis(RA) and should be used inconjunction with other clinicalinformation. Autoantibody levelsrepresent one parameter in a multi-criteria diagnostic process,encompassing both clinical andlaboratory-based assessments.	The DIASTATT™ Anti-CCP test is asemi-quantitative/qualitativeenzyme-linked immunosorbent assay(ELISA) for the detection of the IgGclass of autoantibodies specific tocyclic citrullinated peptide (CCP) inhuman serum or plasma. The test isintended to aid in the diagnosis ofRheumatoid Arthritis (RA) and is notdefinitive in isolation.Autoantibody levels represent oneparameter in a multi-criteriondiagnostic process, encompassingboth clinical and laboratory-basedassessments.
Antigen Usedon the SolidPhase	Synthetic cyclic citrullinatedpeptide, second generation	Same
DetectingAntibody	Murine monoclonal antibodyconjugated to AlkalinePhosphatase	Same
Controls	Negative and positive kit controls	Same
Interference	Rheumatoid factor (up to 200IU/mL does not interfere with anti-CCP antibody results.	Same
DIFFERENCES
Item	Device	Predicate
	IMMULITE® 2000 Anti-CCPIgG Assay	DIASTATT™ Anti-CyclicCitrullinated Peptide (anti-CCP)ELISA
Assay format	Solid-phase, two-cycle sequentialchemiluminescent immunometricassay	Enzyme-linked immunosorbentassay (ELISA)
Substrate	Chemiluminescent	Mg2+, phenolphthaleinmonophosphate (PMP)
Sample size	5 µL(20 µL of the prediluted sample)	10 µL(100 µL of the prediluted sample)
SampleDilution	1:40	1:100
Sample Types	Serum or plasma (EDTA, lithiumheparin)	Serum or plasma (EDTA, lithiumheparin, sodium citrate)
Incubation	30 minutes, 30 minutes, 5 minutes	60 minutes, 30 minutes, 30 minutes
LoD	1.50 U/mL	0.05 U/mL
ExpectedValues(apparentlyhealthydonors)	Median: <1.50 U/mLRange: <1.50 U/mL	Mean : 0.63 ± 0.419U/mLRange: 0.05 - 3.8U/mL
ReportableRange	1.50 - 200 U/mL	0.05 - 100U/mL
Assay Cut-off	≥4.00 U/mL = Reactive	>5 U/mL = Positive
Interference	Bilirubin (up to 0.2 mg/L),hemoglobin (up to 500 mg/dL),and triglycerides (up to 1.5 g/dL)do not interfere with anti-CCPantibody results.Total protein not assessed.	Bilirubin (up to 0.2 mg/L),hemoglobin (up to 400 mg/dL), andintralipid (up to 15 mg/mL) do notinterfere with anti-CCP antibodyresults.Total Protein up to 120 mg/mL doesnot interfere with anti-CCP antibodyresults.

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K. Standard/Guidance Documents Referenced:

CLSI EP5-A2	Evaluation of Precision Performance of Quantitative Measurement Methods
CLSI EP6-A	Evaluation of the Linearity of Quantitative Measurement
CLSI EP17-A	Protocols for Determination of Limits of Detection and Limits of Quantitation
CEN 13640	Stability Testing of In Vitro Diagnostic Reagents

L. Test Principle:

The IMMULITE® 2000 Anti-CCP IgG assay is a solid phase, two-cycle sequential chemiluminescent immunometric assay. In the first cycle, the patient sample and the buffer are incubated together with the coated bead for 30 minutes. During this time, human IgG in the sample binds to CCP on the bead. Unbound sample is then removed by centrifugal washes. In the second cycle, the enzyme conjugated monoclonal murine antihuman IgG is added to the original reaction tube for additional 30 minutes incubation. The enzyme conjugated monoclonal wurine anti-human IgG antibody binds to immobilized anti-CCP IgG on the bead. The unbound enzyme conjugate is removed by centrifugal washes. Finally, chemiluminescent substrate is added to the reaction tube containing the bead and the signal is generated in proportion to the bound enzyme.

M. Performance Characteristics

1. Analytical Performance:
  a. Precision/Reproducibility:

Precision was evaluated using a protocol based on CLSI document EP05-A2, Evaluation of Precision Performance of Quantitative Measurement Methods. Eleven samples were tested over a period of 20 days, using 2 IMMULITE® 2000 instruments with 2 reagent lots each, 2 runs per day and 2 replicates per sample. The work list consisted of autisth control and 2 sets of six-member precision panels: one serum donor panel and one lithium heparin plasma donor panel. Precision panels were prepared by pooling the samples and spiking to the desired concentration with a stock Anti-CCP reactive sample. The sample concentrations are detailed in the table below:

Sample	Target Concentration (U/mL)
LPIC2.101 Positive control	46.0
SERP2 serum donor panel	2.96
SERP3 serum donor panel	4.8
SERP4 serum donor panel	9.0
SERP5 serum donor panel	37.4
SERP6 serum donor panel	135.5
PLASP2 Li Heparin donor panel	2.59
PLASP3 Li Heparin donor panel	4.9
PLASP4 Li Heparin donor panel	9.0
PLASP5 Li Heparin donor panel	37.4

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Sample	Target Concentration (U/mL)
PLASP6 Li Heparin donor panel	135.5

Precision data is summarized in the table below:

20-DayImprecisionN=320		Between Lot		Between Day		Between Run		Within Run		Total
Sample	MeanU/mL	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
LPIC2	47.4	0.18	0.4	0.00	0.0	0.90	1.9	1.84	3.9	2.05	4.3
SERP2	2.1	0.20	9.3	0.15	6.9	0.15	6.8	0.20	9.4	0.29	13.6
SERP3	4.3	0.08	1.8	0.13	3.1	0.17	3.9	0.28	6.5	0.35	8.2
SERP4	8.5	0.00	0.0	0.18	2.1	0.24	2.9	0.34	4.0	0.46	5.4
SERP5	37.6	0.66	1.8	0.96	2.6	1.12	3.0	1.54	4.1	2.13	5.7
SERP6	144.0	2.29	1.6	1.25	0.9	4.17	2.9	5.45	3.8	6.98	4.8
PLASP2	2.4	0.16	6.7	0.17	6.8	0.17	7.1	0.20	8.2	0.31	12.8
PLASP3	4.5	0.18	3.8	0.17	3.6	0.20	4.5	0.29	6.4	0.39	8.6
PLASP4	8.4	0.00	0.0	0.05	0.6	0.31	3.7	0.35	4.1	0.47	5.6
PLASP5	38.5	0.11	0.3	0.54	1.4	1.04	2.7	1.57	4.0	1.96	5.0
PLASP6	141.5	1.06	0.7	2.23	1.6	3.87	2.7	5.52	3.9	7.11	5.0

Reproducibility was evaluated using two different reagent lots at three external testing sites, using serum donor panels from the precision study. The protocol was run over 10 days, 2 runs per day, with 4 replicates per run for the sample pools and control material r

The results for reproducibility, pooled across 3 sites, are presented in the table below.

Sample ID	N	Mean(U/mL)	Within Run		Between Run		Between Day		Between Site		Total Within-Device		Total Within-Lot
			SDIndex	CV(%)	SDIndex	CV(%)	SDIndex	CV(%)	SDIndex	CV(%)	SDIndex	CV(%)	SDIndex	CV(%)
Reagent Lot 201
SERP2	244	1.94	0.20	10.1	0.06	3.2	0.11	6.0	0.17	8.7	0.23	12.0	0.29	14.8
SERP3	244	3.98	0.22	5.6	0.12	2.9	0.11	3.0	0.14	3.6	0.27	6.9	0.31	7.8
SERP4	244	8.08	0.37	4.5	0.07	0.8	0.17	2.0	0.18	2.3	0.41	5.1	0.45	5.6
SERP5	243	36.07	1.48	4.1	0.50	1.4	0.61	2.0	0.84	2.3	1.68	4.6	1.88	5.2
SERP6	244	139.18	5.11	3.7	2.84	2.0	1.25	1.0	3.79	2.7	5.98	4.3	7.08	5.1

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Sample ID	N	Mean(U/mL)	Within Run		Between Run		Between Day		Between Site		Total Within-Device		Total Within-Lot
			SD Index	CV (%)	SD Index	CV (%)	SD Index	CV (%)	SD Index	CV (%)	SD Index	CV (%)	SD Index	CV (%)
LPIC2	246	45.66	1.97	4.3	0.56	1.2	0.83	2.0	1.55	3.4	2.21	4.8	2.70	5.9
Reagent Lot 202
SERP2	228	1.61	0.19	11.7	0.04	2.2	0.13	8.0	0.16	10	0.23	14.4	0.28	17.6
SERP3	240	3.72	0.25	6.7	0.09	2.4	0.21	6.0	0.15	4.1	0.34	9.1	0.37	10
SERP4	240	8.02	0.41	5.2	0.22	2.7	0.16	2.0	0.27	3.4	0.50	6.2	0.56	7.0
SERP5	240	36.79	1.36	3.7	0.86	2.3	0.89	2.0	1.10	3.0	1.84	5.0	2.14	5.8
SERP6	240	139.61	6.27	4.5	2.46	1.8	4.81	3.0	4.81	3.4	8.28	5.9	9.58	6.9
LPIC2	240	43.54	2.49	5.7	0	0	1.61	4.0	1.65	3.8	2.97	6.8	3.39	7.8

b. Linearity/assay Reportable Range: The Anti-CCP Linearity study was performed according to the CLSI document EP6-A; Evaluation of the Linearity of Quantitative Measurement Methods. A dilution series was prepared by combining a reactive serum pool and a non-reactive serum pool in different ratios to produce 11 dilutions covering the assay measuring range (1.50 U/mL to 200 U/mL). The dilutions were run in triplicate on one IMMULITE® 2000 instrument. Results are shown below. Since this assay has no reference material or method and is traceable to internal standard, the recovery is defined as the difference between the observed dose and the fitted values of the linear equation.

Dose Recovery:
Sample	ExpectedDose(U/mL)	MeanObservedDose(U/mL)	LinearFittedDose(U/mL)	Mean% Recovery
P3	181.51	182.93	183.00	100.0%
P4	151.38	158.52	152.70	103.8%
P5	121.26	127.68	122.40	104.3%
P6	91.13	93.00	92.09	101.0%
P7	61.00	61.91	61.79	100.2%
P8	30.87	28.32	31.49	89.9%
P9	15.80	17.01	16.34	104.1%
P10	12.07	11.89	12.58	94.5%
P11	8.33	8.58	8.82	97.3%
P12	4.43	4.48	4.89	91.5%
P13	2.64	2.40	3.10	77.4%

c. Traceability, Stability, Expected Values (controls, calibrators, methods):

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Traceability - There is no recognized standard reference material for Anti-CCP. The IMMULITE® 2000 Anti-CCP IgG assay is traceable to an internal standard and manufactured using qualified materials and measurement procedures.

Stability - Freshly opened and prepared IMMULITE® 2000 Anti-CCP IgG adjustors and controls showed 30 days open vial stability when stored at 2-8°C, or 6 months (aliquotted) when stored at -20°C. The IMMULITE® 2000 Anți-CCP IgG reagent wedge is stable at 2-8°C until the expiration date on the label.

Adjustors and Controls are prepared in house and arbitrary units are assigned during the development process. Calibrator and Control values are summarized in the tables below. The listed values are approximate values, as the values are lot specific.

Adjustor	Dose Range U/mL
Low	2.0 - 5.0
High	40 - 60

Control	Target* U/mL
Low	<1.5
High	44

d. Detection Limit:

Limit of Blank (LoB): An Anti-CCP nonreactive donor sample was used as a blank and was analyzed on three IMMULITE® 2000 instruments over 5 days. The sample was tested twice daily using two lots of reagent with two replicate per run. The LoB result was determined by applying a nonparametric principle based on ran! 1 ho Hored value using the following equation:

LoB = [Ng(p/100)+0.5]; where p=(100-α) = 95 and NB = number of blank measurements = 732

The LoB was the average of the ranked results at positions 695 and 696. The LoB was determined to be 0.26 U/mL.

Limit of Detection (LoD): Five Anti-CCP serum samples (mean concentrations ranging from 0.26 to 2.5 U/mL) were assayed in replicates of 6 using 2 reagent kit lots run for 5 days with 2 runs per day. Two instrument and 2 operators generated 960 observations.

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LoD = LoB +median -5th percentile.

LoD = 0.26 + 1.4 -0.2 = 1.46 U/mL.

e. Analytical Specificity:
Endogenous Interferents: A study was conducted to assess the effect of several endogenous interferents on the IMMULITE®2000 Anti-CCP IgG assay. Testing consisted of the addition of human serum albumin, triglycerides, hemogly bin, bilirubin (conjugated and unconjugated), and rheumatoid factor into the samples. One control sample and five (5) different sample pools with differing Anti-CCP concentrations (range 2.0 U/mL to 205 U/mL) were used. Each potentially interfering substance was spiked separately into the sample pools. Results are shown in the table below.

IMMULITE 2000 Anti-CCP:Summary of EndogenousInterferents in Anti-CCPSamples	% Difference:Control Dose/ Test Dose:Lot 201	% Difference:Control Dose/ Test Dose:Lot 202	Mean % InterferenceLot 201 & Lot202
Human Serum Albumin (12g/dL)	1.2	6.9	4.1
Triglycerides (1.5 g/dL)	1.8	-1.4	0.2
Hemoglobin (500 mg/dL)	4.5	2.1	3.3
Conjugated Bilirubin (0.2mg/mL)	-1.6	-2.0	-1.8
Unconjugated Bilirubin (0.2mg/mL)	-3.6	-2.9	-3.3
Rheumatoid Factor (200IU/mL)	0.8	-4.1	-1.7

Less than 10% mean interference was found with all endogenous interfering substance as follows: Human Serum Albumin (up to 12g/dL); Triglycerides (up to 1.5 g/dl); Hemoglobin (500 mg/dL); Bilirubin: Conjugated and Unconjugated (up to 0.2 mg/mL each); and Rheumatoid Factor (up to 200 IU/mL). However, individual samples occasionally showed > ± 10% interference.

Hook Effect: Not applicable. The IMMULITE® 2000 Anti-CCP assay is a 2-cycle assay with sample containing anti-CCP antibodies first binding to the CCP pertide immobilized on the polystyrene capture bead. Excess unbound antibody is pephued by washing prior to the second cycle reagent incubation. During the second chive, an anti-human IgG alkaline phosphatase conjugate binds in turn to the anti-CCP antibody bound to the bead.

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Assay response is positively related to the amount of anti-CCP antibody bound to the capture bead up to the point where the CCP determinants are saturated with antibody, at which point the response plateaus. Any unbound anti-CCP beyond the reportable range of the assay and in excess of saturation is removed by washing prior to contact with the reagent conjugate and there is no risk that a hook effect will be observed.

f. Assay cut-off:

A high Anti-CCP IgG positive defibrinated plasma unit was arbitrarily assigned with a value of 1000 U/mL. A lot of reference calibrators was prepared by diluting the positive unit with negative normal human serum. The reference calibrators were value-assigned by the dilution factor in relationship to the neat values of 1000 U/mL.

A total of 388 samples consisting of 212 apparently healthy samples, 106 RA positive samples, 70 anti-CCP positive samples, as determined by a 3rd party method, were tested with IMMULITE 2000 Anti-CCP Kit and the DIASTAT Anti-CCP ELISA.

The cutoff of the IMMULITE® 2000 Anti-CCP IgG assay was determined with positive and negative patient samples by a ROC analysis, with a balanced consideration of sensitivity and specificity. A result greater than or equal to 4U/mL indicates that anti-CCP IgG antibodies were detected in the sample. A result of less than 4U/mL indicates that anti-CCP IgG antibodies were not detected in the sample.

2. Comparison Studies

a. Method Comparison with predicate device:

Method Comparison: The IMMULITE® 2000 Anti-CCP IgG assay was compared to the predicate Axis-Shield DIASTAT™ Anti-CCP assay using two lots of reagents. Each sample was tested in singlicate using 3 IMMULITE® 2000 instruments. For reagent lot 1, 255 unaltered patient serum samples were tested, 229 of which were determined to be reactive and 26 nonreactive using the DIASTATTM assay. For reagent lot 2, 256 unaltered patient serum samples were tested, 230 of which were determined to be reactive and 26 nonreactive using the DIASTAT™ assay. Results are shown in the tables below.

Reagent lot 1:

IMMULITE 2000 Anti-CCP IgG

DIASTAT	Reactive	Non-Reactive	Percent PositiveAgreement	Percent NegativeAgreement
Positive	199	30	86.9%
Negative	14	12		46.2%

Total Agreement: 82.2%

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Reagent lot 2:

IMMULITE 2000 Anti-CCP IgG

DIASTAT	Reactive	Non-Reactive	Percent PositiveAgreement	Percent NegativeAgreement
Positive	198	32	86.1%
Negative	15	11		42.3%

Total Agreement: 81.6%

b. Matrix Comparison:

Thirty-nine matched serum and plasma samples were collected in the following anticoagulant tubes: serum clot tube, Lithium Heparin plasma tube, serum separator tube (SST), and EDTA plasma tube. Twenty-one of the 39 samples were spiked to achieve Anti-CCP levels across the assay measuring range. Data was analyzed using Deming regression plots.

SERUM vs.	CorrelationCoefficient	Slope	Intercept	Means
Lithium Heparin	1.000	1.020	-0.46	52.9 U/mL
SST Serum	1.000	1.020	-0.52	53.0 U/mL
EDTA Plasma	1.000	1.010	-0.31	52.2 U/mL
				Serum: 52.3 U/mL

3. Clinical Studies:

a. Clinical Sensitivity and Specificity

Clinical Performance Study: The assay was performed on the IMMULITE® 2000 to assess the clinical sensitivity and specificity of well-characterized samples.

A total of 1512 patient serum samples were collected for the study. 1048 samples were rheumatoid arthritis (RA) positive and the remaining 464 samples were from not-RA patients with potentially cross-reactive diseases. RA positive patients were classified according to the ACR criteria.

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IMMULITE 2000 Anti-CCP IgG

Clinical Status	Reactive	Non-Reactive	Sensitivity	Specificity
Positive	667	381
Negative	14	450	63.6%	97.0%

b. Other clinical supportive data (when a. and b. are not applicable): Not applicable.
1. Clinical Cut-off: Same as assay cut-off.
1. Expected Values/Reference Range:

A total of 200 serum samples from presumed healthy male and female donors were analyzed using the IMMULITE® 2000 Anti-CCP IgG assay. The results from this study suggest a median of <1.5 U/mL, and a 99th percentile of 4.06 U/mL. Expected walves in the normal population should be negative. Each laboratory should establish a marks i range appropriate to their patient populations and clinical practice.

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IMMULITE® 2000 Anti-CCP IgG CVM

510(k) Summary as Required by 21 CFR 807.92

A. 510(k) NumberB. Purpose forSubmission	New device
C. Measurand	Anti-cyclic citrullinated peptide (CCP) antibodies
D. Type of test	Calibration verification material for IMMULITE Anti-CCP IgG assay
E. Applicant:	Siemens Healthcare Diagnostics Inc.511 Benedict AvenueTarrytown, NY 10591
F. Proprietary andEstablished Name:	IMMULITE® 2000 Anti-CCP IgG Calibration Verification Material
G. RegulatoryInformation:
1. Regulation section:	21 CFR 862.1660
2. Classification:	Class I (reserved)
3. Products Codes:	JJX - Single Analyte Control (assayed and unassayed)
4. Panel:	Clinical Chemistry (75)
H. Intended Use:	The IMMULITE 2000 Anti-CCP IgG Calibration VerificationMaterial is for in vitro diagnostic use, as a control for calibrationverification of the IMMULITE® 2000 Anti-CCP IgG assay on theIMMULITE 2000 system.
I. Device Description:	The IMMULITE Anti-CCP IgG Calibration Verification Materialconsists of one set of four vials, containing low, intermediate and highlevels of lyophilized human serum with IgG reactive to cycliccitrullinated peptide (CCP), in buffer with preservative, plus ananti-CCP-free sample.
	Source materials derived from human blood were tested and foundnonreactive for syphilis; for antibodies to HIV 1 and 2; for hepatitis Bsurface antigen; and for antibodies to hepatitis C.
J. SubstantialEquivalenceInformation:	Predicate device name: Elecsys Anti-CCP CalCheck510(k) number: K091601

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Comparison with predicate:

A comparison of the device features, intended use, and other information demonstrates that the IMMULITE ® 2000 Anti-CCP IgG Calibration Verification Material is substantially equivalent to the predicate device, Roche Elecsys Anti-CCP CalCheck, as summarized in the following tables.

SIMILARITIES
Item	Device	Predicate
	IMMULITE Anti-CCP IgG CVM	Elecsys Anti-CCP CalCheck
Intended Use	For in vitro diagnostic use, as a control for calibration verification of Anti-CCP IgG assay.	Same
Format	Lyophilized	Same
Matrix	Human serum	Same
Handling	Reconstitute with distilled or deionized water	Same

DIFFERENCES
Item	Device	Predicate
	IMMULITE Anti-CCP IgGCVM	Elecsys Anti-CCP CalCheck
Analyzer	For use on IMMULITE 2000analyzer	For use on Elecsys 2010,MODULAR ANALYTICS E170,cobas e411 and cobas e601analyzers
CVM Levels	4	3

K. Standard/Guidance Documents Referenced:

Guidance for Industry and FDA Staff Assayed and Unassayed Quality Control Material .

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L. Test Principle:

. Not Applicable

M. Performance Characteristics

6. Analytical Performance:

a. Precision/Reproducibility: Not Applicable
b. Linearity/assay reportable range: Not Applicable
c. Traceability, Stability, Expected values:

Traceability - Since there is no recognized standard reference material for Anti-CCP, the IMMULITE 2000 Anti-CCP IgG assay is traceable to an internal standard and manufactured using qualified materials and measurement procedures. The IMMULITE Anti-CCP Calibration Verification Material (CVM) is traceable to this standard.

Stability - Open vial stability - Freshly opened and reconstituted IMMULITE 2000 Anti-CCP CVMs showed 30 days open vial stability when stored at 2-8°C. Unopened stability is indicated by expiration date on the label when stored at 2-8°C.

Value Assignment: The Anti-CCP reference calibrator values were assigned by using an approved internal standard. This reference standard lot is used to assign Anti-CCP IgG CVMs. Quality control is then performed by calculating the recovery of controls using the reference calibration verification material.

Each CVM level below was run in duplicate on 3 different reagent kit lots (Lot 201, 202 and 211) and 14-15 runs per lot for a total of 86 replicates. Five instruments were used to carry out all the runs. The analyte values were calculated based on the recovered values for each run independently. The average analyte recovered for each CVM level determined the value assigned to the Target Mean.

The Guideline Range (95% confidence interval) for each CVM level was established based on the Target Mean and ± 2 Standard Deviations (SD).

Level	Catalog andLot number	Target Mean(U/mL)*	SD	GuidelineRange(U/mL)	TotalCV%	SpecCV%
1	L2PICVM1 Dxxx	0.00		≤ 1.50	NA**	NA
2	L2PICVM2 Dxxx	6.19	0.41	5.377.00	6.6%	10%
3	L2PICVM3 Dxxx	103	8.85	85.3121	8.6%	10%
4	L2PICVM4 Dxxx	209	21.09	167252	10%	10%

note that CVM value assignment is lot-specific

** NA = Not Applicable

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d. Detection limit: Not Applicable
e. Analytical Specificity: Not Applicable
Assay cut-off: Not Applicable · f.

7. Comparison Studies

Method Comparison with predicate device: Not Applicable a.
Matrix Comparison: Not Applicable b.

8. Clinical Studies:

a. Clinical Sensitivity and Specificity: Not Applicable
b. Other clinical supportive data (when a. and b. are not applicable): Not Applicable
1. Clinical Cut-off. Not Applicable
1. Expected Values/Reference Range: See Value Assignment above.

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Image /page/16/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES • USA" arranged around the perimeter. Inside the circle is a stylized design featuring a symbol that resembles a human figure embracing or protecting another figure, representing the department's mission of protecting the health of all Americans and providing essential human services.

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

April 4, 2013

Siemens Healthcare Diagnostics c/o Susan Brocchi Sr. Regulatory Affairs Specialist 511 Benedict Avenue Tarrytown, NY 10591

Re: K121576

Trade/Device Name: IMMULITE 2000 Anti-CCP IgG Assay and IMMULITE 2000 Anti-CCP IgG Calibration Verification Material Regulation Number: 21 §CFR 866.5775 Regulation Name: Rheumatoid factor immunological test system Regulatory Class: Class II Product Code: NHX, JIT Dated: March 29, 2013 Received: April 1, 2013

Dear Ms. Brocchi:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food. Drug. and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you; however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21

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Page 2 - Ms. Susan Brocchi

CFR Part 807); labeling (21 CFR Part 801); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801), please contact the Division of Small Manufacturers, International and Consumer Assistance at its tollfree number (800) 638-2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm. Also, please note the regulation entitled. "Misbranding by reference to premarket notification" (21CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to

http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

Maria M. Chan -S

Maria M. Chan, Ph.D. Director Division of Immunology and Hematology Devices Office of In Vitro Diagnostics and Radiological Health (OIR) Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known): kl21576

Device Name: IMMULITE® 2000 Anti-CCP IgG Assay and Calibration Verification Material

Indications for Use: The IMMULITE 2000 Anti-CCP IgG assay is an in vitro diagnostic immunoassay for the semi-quantitative determination of the IgG class of autoantibodies specific to cyclic citrullinated peptide (CCP) in human serum (including Serum Separator Tubes) or plasma (EDTA or lithium heparin) on the IMMULITE 2000 system. Detection of anti-CCP antibodies is 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 multi-criteria diagnostic process, encompassing both clinical and laboratory-based assessments.

Prescription Use X (21 CFR Part 801 Subpart D) Over the Counter Use (21 CFR Part 801 Subpart C)

(PLEASE DO NOT WRITE BELOW THIS LINE; CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH, Office of In Vitro Diagnostics and Radiological Health (OIR)

And/Or

Maria M. Chan -S

Division Sign-Off Office of In Vitro Diagnostics and Radiological Health

510(k) K121576

Regulation Number and Section

§ 866.5775 Rheumatoid factor immunological test system.

(a)
Identification. A rheumatoid factor immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the rheumatoid factor (antibodies to immunoglobulins) in serum, other body fluids, and tissues. Measurement of rheumatoid factor may aid in the diagnosis of rheumatoid arthritis.(b)
Classification. Class II (performance standards).