The EUROIMMUN Anti-PLA2R ELISA (IgG) test kit is intended for the qualitative or semiquantitative determination of IgG class autoantibodies against phospholipase A2 receptor (PLA2R) in human serum. It is used as an aid in the diagnosis of primary membranous glomerulonephritis (pMGN), in conjunction with other laboratory and clinical findings.

Device Description

The EUROIMMUN Anti-PLA2R ELISA (IgG) consists of a microwell ELISA plate coated with PLA2R antigen, 5 calibrators, positive and negative control, peroxidase-labelled anti-human IgG conjugate, sample buffer, wash buffer concentrate, TMB chromogen/substrate solution and stop solution.

AI/ML Overview

The acceptance criteria and study proving the device meets them are detailed below for the EUROIMMUN Anti-PLA2R ELISA (IgG).

1. Table of Acceptance Criteria and Reported Device Performance

Performance Characteristic	Acceptance Criteria	Reported Device Performance
Reproducibility
Intra-Assay CV	< 12% for positive and borderline samples; all qualitative results for negative samples to be negative; all qualitative results (positive, borderline, negative) of ratio-based samples to be in line with expected result.	RU/ml: CVs range from 2.6% to 10.9%. Ratio: All qualitative results in line with expected (e.g., 0% positive for negative, 100% positive for positive).
Inter-Assay CV	< 12% for positive and borderline samples; all qualitative results for negative samples to be negative; all qualitative results (positive, borderline, negative) of ratio-based samples to be in line with expected result.	RU/ml: CVs range from 4.2% to 10.3%. Ratio: All qualitative results in line with expected (e.g., 100% negative for negative, 100% positive for positive, 73% borderline for a borderline sample).
Repeatability (Total Imprecision)	< 12% for positive and borderline samples; all qualitative results for negative samples to be negative; all qualitative results (positive, borderline, negative) of ratio-based samples to be in line with expected result.	RU/ml: Total CVs range from 3.6% to 14.8% (Sample 1 above 12%). Ratio: All qualitative results largely in line with expected (e.g., 100% negative for negative, 100% positive for positive, 97-99% borderline for borderline samples).
Lot-to-Lot Reproducibility	< 12% for positive and borderline samples; all qualitative results for negative samples to be negative; all qualitative results (positive, borderline, negative) of ratio-based samples to be in line with expected result.	RU/ml: Total CVs range from 4.0% to 11.2% (Sample 1 is 11.2%). Ratio: All qualitative results largely in line with expected (e.g., 100% negative for negative, 100% positive for positive, 72-94% borderline for borderline samples).
Linearity/Assay Reportable Range	Sufficiently linear from 2 to 1500 RU/mL.	The assay was shown to be sufficiently linear from 2 to 1500 RU/mL.
Limit of Detection (LoD)	LoD should be approximately 2.2 RU/ml (or lower to functional sensitivity).	LoD was found to be 2.2 RU/ml.
Limit of Blank (LoB)	Not explicitly stated an acceptance criterion, but investigated.	LoB was found to be 1.8 RU/ml.
Limit of Quantitation (LoQ)/Functional Sensitivity	LoQ = LoD, or approximately 1.4 RU/ml if using functional sensitivity.	Functional sensitivity was approx. 1.4 RU/ml. LoQ = LoD = 2.2 RU/ml.
Analytical Specificity (Cross-reactivity)	No significant cross-reactivity with tested disease states.	All 65 sera (thyroiditis, SLE, Sjögren's, SSc, RA, cANCA, pANCA, GBM, HBsAg) were negative, indicating no cross-reactivity.
Analytical Specificity (Interference)	Individual recovery within 70-130%; mean recovery within 85-115%.	No significant interference for hemoglobin (up to 1000 mg/dl), triglyceride (up to 2000 mg/dl), and bilirubin (up to 40 mg/dl).
Method Comparison (Predicate Device)	Not explicitly stated with numerical values, but implied substantial agreement.	RU/mL: Positive Agreement (borderline as negative): 86.79% (95% CI: 81.5%-91.0%). Negative Agreement: 99.71% (95% CI: 98.4%-100.0%).RU/mL: Positive Agreement (borderline as positive): 89.62% (95% CI: 84.7%-93.4%). Negative Agreement: 99.71% (95% CI: 98.4%-100.0%).Ratio: Positive Agreement (borderline as negative): 86.79% (95% CI: 81.5%-91.0%). Negative Agreement: 99.71% (95% CI: 98.4%-100.0%).Ratio: Positive Agreement (borderline as positive): 92.45% (95% CI: 88.0%-95.6%). Negative Agreement: 99.71% (95% CI: 98.4%-100.0%).
Clinical Sensitivity	Within the expected range of approximately 70% as reported in scientific literature.	66.9% (95% C.I.: 61.0 - 72.4%) for pMGN, using 5-point calibrated analysis and 20 RU/ml cut-off (borderline samples counted as negative). 68.7% (95% C.I.: 62.9 - 74.2%) for pMGN (borderline samples counted as positive).
Clinical Specificity	Not explicitly quantified, but demonstrated high specificity against various control groups.	99.6% (95% C.I.: 98.1 - 100.0%) against all control groups combined (borderline samples counted as negative or positive).

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

Reproducibility (Intra-Assay): 20 determinations for RU/ml based results, 20 determinations for Ratio based results.
Reproducibility (Inter-Assay): 30 determinations for RU/ml based results, 30 determinations for Ratio based results.
Repeatability (Total Imprecision): 150 determinations per sample for RU/ml and Ratio based results.
Reproducibility (Lot-to-Lot): 18 determinations per sample for RU/ml and Ratio based results.
Linearity/Assay Reportable Range: Five sets of 11-step-wise dilutions (totaling 55 samples).
Limit of Blank (LoB), Limit of Detection (LoD), Limit of Quantitation (LoQ): Investigated following CLSI standard EP17-A.
Analytical Specificity (Cross-reactivity): Panel of 65 clinically characterized sera.
Analytical Specificity (Interference): Sera with anti-PLA2R concentrations spiked with potential interfering substances.
Method Comparison and Clinical Study:
- Test Set Size: 560 samples in total; 275 from pMGN patients and 285 from control groups.
- Data Provenance: The document states "Clinical studies were performed in cooperation with different sites." While specific countries are not mentioned, it implies multiple clinical sites. The studies are prospective in nature in the sense that samples were collected for analysis by the new device against established clinical diagnoses. The clinical diagnoses were based on renal biopsy and clinical/laboratory criteria, and samples were drawn within 8 weeks after biopsy before treatment.

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

The document does not explicitly state the number of experts or their specific qualifications (e.g., "radiologist with 10 years of experience") used to establish the ground truth for the clinical study samples.
However, the clinical study section states: "pMGN diagnosis was based on renal biopsy and was considered to be idiopathic/primary when no secondary cause of MN was suspected on the basis of clinical and laboratory criteria." This implies a standard clinical diagnostic process involving medical professionals (e.g., nephrologists, pathologists) who interpret biopsies and clinical data.

4. Adjudication Method for the Test Set

The document does not describe an explicit adjudication method for the clinical study's ground truth beyond stating that pMGN diagnosis was "based on renal biopsy and was considered to be idiopathic/primary when no secondary cause of MN was suspected on the basis of clinical and laboratory criteria." This suggests that the clinical diagnosis, informed by biopsy and other findings, served as the ground truth. There is no mention of a process like "2+1" or "3+1" for reaching a consensus diagnosis for individual cases.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance

This information is not applicable to the EUROIMMUN Anti-PLA2R ELISA (IgG) device. This is an in vitro diagnostic (IVD) assay, not an AI-powered image analysis or diagnostic support system that would involve human readers and AI assistance. The performance is measured directly through laboratory analysis of patient samples.

6. If a Standalone (i.e. algorithm only without human-in-the loop performance) was done

This is an in vitro diagnostic (IVD) assay (ELISA), not an algorithm or AI system. The performance characteristics (sensitivity, specificity, reproducibility, etc.) are inherent to the assay and its protocol when performed in a laboratory setting. Therefore, a "standalone algorithm performance" study is not relevant in this context. The results are generated by the assay itself, following the prescribed procedure, and then interpreted based on established cut-off values.

7. The Type of Ground Truth Used

Clinical Study: The ground truth for the clinical study samples was based on clinical diagnosis, primarily established by:
- Renal biopsy for confirmation of primary membranous glomerulonephritis (pMGN).
- Clinical and laboratory findings to rule out secondary causes of membranous nephropathy.
Analytical Studies (e.g., Reproducibility, Linearity, Cross-reactivity, Interference): The ground truth for these studies was based on:
- Known concentrations or expected relative values of analytes in prepared samples or controls.
- Known clinical characteristics of patient panels (e.g., positive for thyroiditis, negative for anti-PLA2R antibodies, etc.).

8. The Sample Size for the Training Set

This is an in vitro diagnostic (IVD) assay, not a machine learning or AI model that requires a distinct "training set" in the traditional sense. The device's performance characteristics are established through various analytical and clinical studies, but there isn't a "training phase" to optimize an algorithm with a specific dataset.

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

As explained above, this device is not an AI/ML algorithm requiring a training set. The "ground truth" equivalent for the development and validation of an IVD assay involves:
- Reference materials/calibrators: Precisely prepared solutions with known concentrations of the analyte.
- Clinically characterized samples: Sera from patients with confirmed diagnoses (e.g., pMGN by biopsy, specific autoimmune diseases) and healthy controls, used to determine the assay's clinical performance. These samples are analogous to validation data for an algorithm.

Summary

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K132195

JUN 2 7 2014

PREMARKET NOTIFICATION 510(K) SAFETY AND EFFECTIVENESS SUMMARY

(As required by 21 CFR § 807.92)

A. 510(k)Number: K132195
B. Purpose for Submission: New Device
C. Measurand: Anti-PLA2R Autoantibodies
D. Type of Test: Qualitative or Semi-Quantitative Enzyme Immunoassay
E. Applicant: EUROIMMUN US INC.
F. Proprietary and Established Names: EUROIMMUN Anti-PLA2R ELISA (IgG)

G. Regulatory Information:

1. Regulation: 21 CFR 866.5780 - Anti-phospholipase A2 receptor immunological test system
1. Classification: Class II
1. Product code: PGV - Anti-phospholipase A2 receptor
1. Panel: Immunology

H. Intended Use:

1. Intended Use(s):
  The EUROIMMUN Anti-PLA2R ELISA (IgG) test kit is intended for the qualitative or semiquantitative determination of IgG class autoantibodies against phospholipase A2 receptor (PLA2R) in human serum. It is used as an aid in the diagnosis of primary membranous glomerulonephritis (pMGN), in conjunction with other laboratory and clinical findings.
Indication(s) for Use: 2. Same as Intended Use.
1. Special Conditions for the Use Statement(s): For Prescription Use Only.
1. Special Instrument Requirements: Microwell plate reader capable of measuring OD at 450nm for dual wavelength readings.

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EUROIMM

I. Device Description:

J. Substantial Equivalence Information:

Predicate device name (s): 1. EUROIMMUN Anti-PLA2R IFA (IgG)
1. Predicate 510(k) number(s): K132379
1. Comparison with predicate:

Similarities

Item	New device	Predicate device
Manufacturer	EUROIMMUN AG	Same
Intended use	Detection of IgG antibodies against PLA2R	Same
Sample types	Serum	Same
Controls	2 controls: 1 positive, 1 negative	Same
Reagent preparation	All reagents are ready to use, except for the washbuffer.	Same

Differences

Item	New device	Predicate device
Assay format	Qualitative or semi-quantitative (using either allcalibrators or the cut-off calibrator only)	Qualitative
Antigen	Recombinant PLA2R (type M)	PLA2R transfected cells andcontrol-transfected cells
Reagents	96 well microplate, 5 Calibrators (2, 20, 100, 500and 1500 RU/ml), Conjugate (anti-human IgGlabeled with horseradish peroxidase), Samplebuffer, Wash buffer (10x concentrate), Substratesolution (TMB), Stop solution (0.5 M sulphuricacid), 2 Controls,	BIOCHIP slides, Conjugate(fluorescein-labeled anti-humanIgG), Salt for PBS pH 7.2, Tween20, Embedding medium, Coverglasses, 2 Controls
Sample dilution	1:101	1:10
Procedure	ELISA: Sample incubation with micro-wellantigen coated plate, followed by a wash step,incubation with conjugate, wash step, incubationwith substrate, addition of stop solution,photometric reading	IFA: Sample incubation withtissues/cells, followed by a washstep, incubation with conjugate,wash step, embedding,fluorescence microscopy reading
Reported results	Qualitative, RU/ml or Ratio	Qualitative
Cut-off level	Qualitative: Ratio 1.0Semi-quantitative: 20 RU/ml	1:10 dilution

K. Standard/Guidance Document Referenced (if applicable):

None Referenced.

L. Test Principle:

Patient samples are diluted 1:101 in sample buffer, 100 µl of each diluted patient sample and pre-diluted controls and calibrators are added to the antigen coated microtiter wells and incubated for 30 minutes at room temperature. After incubation the microtiter well strips are washed with wash buffer to remove unbound antibodies and 100 ul of the anti-human IgG enzyme conjugate reagent is added to each microtiter well. After an additional 30-minutes incubation at room temperature, the microtiter wells are again washed 3 times with 300 µl of wash buffer to remove any unbound enzyme conjugate and 100 µl of

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the chromogen substrate is added. The strips are incubated for 15 minutes at room temperature and 100 ul stop solution is added. The microtiter plates are placed in an ELISA reader and read at a wavelength of 450 nm and a reference wavelength of between 620 nm and 650 nm within 30 minutes.

M. Analytical Performance Characteristics (where applicable):

1. Reproducibility
- Intra- and Inter-Assay Reproducibility: a.

Intra- and Inter-Assay coefficients of variation (CV) were determined using samples with values at different points on the calibration curve. The Intra-Assay CVs are based on 20 determinations and the Inter-Assay CVs on 30 determinations performed in 10 different runs on 5 different days (with 3 replicates per run). Tests were performed according to the package insert with the same lot and by the same technician. Acceptance criterium was that the CV's show results below 12% for positive and borderline samples. Acceptance criterium for negative samples was that all qualitative results be negative. Acceptance criterium for the ratio-based results was that all qualitative results (positive, borderline, negative) of the samples be in line with the expected result.

n = 20	Anti-PLA2R ELISA (IgG); RU/ml
	1	2	3	4	5	6	7	8
	Mean Value	2	12	18	26	48	109	782
StDev	0.1	0.5	0.5	0.9	1.5	3.0	33.5	48.7
%CV	10.9	4.2	2.6	3.4	3.1	2.8	4.3	5.7

Intra-Assay Reproducibility

	Anti-PLA2R ELISA (IgG); Ratio
n = 20	1	2	3	4	5	6	7	8
Mean Value	0.1	0.6	0.9	1.3	2.0	4.2	7.5	7.6
Range	0.1 - 0.1	0.5 - 0.6	0.9 - 0.9	1.2 - 1.4	1.9 - 2.1	4.0 - 4.3	7.4 - 7.6	7.5 - 7.7
Expected	neg	neg	bl	pos	pos	pos	pos	pos
% Positive	0%	0%	0%	100%	100%	100%	100%	100%
% Borderline	0%	0%	100%	0%	0%	0%	0%	0%
% Negative	100%	100%	0%	0%	0%	0%	0%	0%

Inter-Assay Reproducibility

n = 30	Anti-PLA2R ELISA (IgG); RU/ml
	1	2	3	4	5	6	7	8
Mean Value	2	12	20	28	51	110	793	884
StDev	1.2	1.0	1.7	1.1	3.2	11.2	81.4	87.5
%CV	---	7.9	8.6	4.2	6.2	10.2	10.3	9.9

n = 30	Anti-PLA2R ELISA (IgG); Ratio
	1	2	3	4	5	6	7	8
Mean Value	0.1	0.6	1.0	1.3	2.2	3.7	7.8	7.9
Range	0.1 - 0.3	0.5 - 0.6	0.8 - 1.1	1.2 - 1.4	1.8 - 2.5	3.0 - 4.3	6.6 - 8.9	7.0 - 8.9
Expected	neg	neg	pos	pos	pos	pos	pos	pos
% Positive	0%	0%	27%	100%	100%	100%	100%	100%
% Borderline	0%	0%	73%	0%	0%	0%	0%	0%
% Negative	100%	100%	0%	0%	0%	0%	0%	0%

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Repeatability & Reproducibility (Total Imprecision) b.

Repeatability
Investigated using samples with values at different points on the calibration curve. Within-run, between-run, between-day and total standard deviations (SD) and coefficients of variation (CV) were calculated based on 150 determinations per sample performed in 6 different runs on 3 different days (with 2 runs per day and 25 replicates per run) according to the package insert with the same lot and by the same technician. Acceptance criterium was that the CV's show results below 12% for positive and borderline samples. Acceptance criterium for negative samples was that all qualitative results be negative. Acceptance criterium for the ratio-based results was that all qualitative results (positive, borderline, negative) of the samples be in line with the expected result.

Repeatability
---------------	--

n = 150		Anti-PLA2R ELISA (IgG); RU/ml
Sample	Mean	Within-run		Between-run		Between-day		Total
		SD	%CV	SD	%CV	SD	%CV	SD	%CV
1	3	0.28	9.2	0.38	12.9	0.64	22.4	0.43	14.8
2	17	0.57	3.3	1.04	6.4	1.25	7.5	0.95	5.7
3	22	0.87	3.9	1.56	7.3	1.08	5.1	1.17	5.4
4	24	0.75	3.2	1.60	6.7	1.51	6.4	1.28	5.4
5	884	69.95	7.9	72.49	8.3	66.51	7.6	69.65	7.9
6	1356	65.65	4.8	33.59	2.5	45.60	3.4	48.28	3.6

n = 150	Anti-PLA2R ELISA (IgG); Ratio
	1	2	3	4	5	6
Mean Value	0.20	0.74	0.91	1.00	7.57	8.20
Range	0.16 - 0.25	0.68 – 0.81	0.76 - 1.27	0.83 – 1.17	6.34 - 8.22	7.45 – 8.72
Expected	neg	bl	bl	pos	pos	pos
% Positive	0%	0%	1%	52%	100%	100%
% Borderline	0%	97%	99%	48%	0%	0%
% Negative	100%	3%	0%	0%	0%	0%

Reproducibility (Lot-to-Lot)

Investigated using samples with values at different points on the calibration curve. Within-run, between-run, between-lot and total SD's and %CV's were calculated based on 18 determinations per sample performed in 3 different runs on 3 different days (with 3 runs per lot and 2 replicates per run) according to the package insert by the same technician. Acceptance criterium was that the CV's show results below 12% for positive and borderline samples. Acceptance criterium for negative samples was that all qualitative results be negative. Acceptance criterium for the ratiobased results was that all qualitative results (positive, borderline, negative) of the samples be in line with the expected result.

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EUROIMMU I II II II II

Reproducibility
n = 18			Anti-PLA2R ELISA (IgG); RU/ml
Sample	Mean		Within-run SD	%CV	Between-run SD	%CV	Between-Batch SD	%CV	Total SD	%CV
1	3		0.45	16.4	0.15	5.4	0.33	11.9	0.31	11.2
2	17		1.19	7.2	0.42	2.5	0.69	4.1	0.76	4.6
3	23		1.46	6.5	0.30	1.3	0.99	4.4	0.91	4.0
4	26		1.78	6.7	0.85	3.2	1.37	5.2	1.33	5.0
5	304		15.71	5.2	6.48	2.1	10.61	3.5	10.93	3.6
6	1260		127.19	10.1	35.80	2.8	33.53	2.7	65.51	5.2
n = 18	Anti-PLA2R ELISA (IgG); Ratio
	1	2	3	4	5	6
Mean Value	0.19	0.77	0.99	1.12	5.62	8.43
Range	0.16-0.21	0.68-0.83	0.91-1.12	1.06 - 1.20	5.29-6.12	7.71-9.04
Expected	neg	neg	bl	pos	pos	pos
% Positive	0%	0%	28%	100%	100%	100%
% Borderline	0%	94%	72%	0%	0%	0%
% Negative	100%	6%	0%	0%	0%	0%

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Linearity/ Assay Reportable Range: C.

Five sets of 11-step-wise dilutions were prepared by mixing low and high analyte samples. The concentrations ranged from a low concentration of 2 RU/mL to high concentrations of 75 RU/mL, 133 RU/mL, 790 RU/mL, 1100 RU/mL, or 1642 RU/mL. The assay was shown to be sufficiently linear from 2 to1500 RU/mL. Results from the two lowest ranges and the range throughout the AMR are shown below.

Image /page/5/Figure/3 description: The image contains two graphs and two tables that show the expected versus measured concentration for two sets of data. The graphs show the mean, linear mean, 2nd order, and 3rd order concentrations. The tables provide statistical information such as R-squared values, coefficients, standard errors, t-values, p-values, and confidence intervals for each order. The equations and R-squared values for each order are also displayed on the graphs.

Image /page/5/Figure/4 description: This image is a graph that shows the relationship between expected concentration and measured concentration. The x-axis represents the expected concentration, ranging from 0.000 to 140.000, while the y-axis represents the measured concentration, ranging from 0.000 to 60.000. The graph includes a trendline with the equation y=1E-05x^3-0.0033x^2+1.1549x-0.812, and the R-squared value for the trendline is 0.9988.

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Image /page/6/Figure/1 description: This image is a graph titled "Expected vs. Measured Concentrations (Set No. 3)". The graph plots the measured concentration on the y-axis against the expected concentration on the x-axis. There are three data sets plotted: Value 1, Value 2, and Mean, each with a corresponding linear trendline. The trendline equations and R-squared values are displayed on the graph, with the mean having a trendline equation of y=0.9595x-25.981 and an R-squared value of 0.9361.

Traceability, Stability, Expected Values (Controls, Calibrators or Methods): d.
A recognized standard or reference material for anti-PLA2R antibodies is not available. The assay is calibrated in relative arbitrary units (RU/ml). Alternatively, results may be given in ratios.

Stability

Stability studies are conducted following the international standard EN 13640:2002: Stability testing of in vitro diagnostic reagents. Three production lots of all kit reagents are tested. Realtime testing at 2-8°C and accelerated testing at 37°C are conducted. The shelf-life stability is 12 months at 2-8°C. Open-vial stability of the kit is 6 months when stored at 2-8°C. The wash buffer was found to be stable for at least 28 days when diluted to working strength.

Controls & Calibrators

The calibrators and controls are derived from human materials. Human originated material is tested and found negative for HBsAg, anti-HCV, anti-HIV-1 and anti-HIV-2, diluted to the appropriate concentration, stabilized and colored.

Calibrators are adjusted to match the required performance criteria in use with the corresponding microtiter strip lot and the corresponding kit controls.

Negative and Positive Controls are included. The positive control is Ready for Use with a 3-4+ fluorescence. Negative control is Ready for Use and is autoantibody negative. EUROIMMUN US INC. recommends using the positive and negative controls undiluted for the screening protocol.

Limit of Blank, Limit of Detection and Limit of Quantitation/Functional Sensitivity: e.
Limit of blank (LoB) and limit of detection (LoD) and limit of quantitation (LoQ)/functional sensitivity (FS) were investigated following CLSI standard EP17-A. The LoB was found to be 1.8 RU/ml & LoD of the Anti-PLA2R ELISA (IgG) was found to be 2.2 RU/ml.

The LoQ was estimated from the functional sensitivity which is defined as the lowest concentration at which the CV is 20%. From the same data LoB and LoD were calculated, the mean concentrations (X-axis) vs. % CVs (Y-axis) were plotted. The functional sensitivity was read from the potential regression line crossing the 20% CV line and was found to be approx. 1.4 RU/ml, which is in the range of the LoD and the lower limit of the measurement range of 2 RU/ml. Following CLSI standard EP17-A , "If this estimate is less than the defined goal for total error, then: LoQ = LoD"; 2.2 RU/ml.

f. Analytical Specificity:

Cross-reactivity: Cross reactivity was investigated using a panel of 65 clinically characterized sera positive for thyreoiditis, systemic lupus erythematosus (SLE), Sjögren's syndrome (SS),

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systemic sclerosis (SSc), rheumatoid arthritis (RA), cANCA, pANCA, GBM and Hepatitis B surface antigen (HBsAg). All 65 sera were negative in the Anti-PLA2R ELISA (IgG), so no cross reactivity was found.

Interference: To investigate the influence from hemoglobin, triglycerides and bilirubin, sera at anti-PLA2R concentrations were spiked with potential interfering substances and incubated with the test system according to the package insert. The recovery in relation to the unspiked sample without interferent was calculated. Acceptance criterium was that the individual recovery be within the range of 70 - 130 % and the mean of recoveries for each interferent be within the range of 85 - 115 %. No significant interference was observed for concentrations of up to 1000 mg/dl for hemoglobin, 2000 mg/dl for triglyceride and 40 mg/dl for bilirubin.

g. Assay Cut-Off:
Qualitative evaluation: Ratio 1.0; <0.7: negative; ≥0.7 to <1.0: borderline; ≥1.0: positive OD of the control or patient sample = Ratio OD of calibrator 2 Semi-quantitative evaluation: 20 RU/ml; <14 RU/mL: negative; ≥14-<20 RU/mL borderline; ≥20 RU/mL positive

Comparison Study(s): 1.

Method comparison with predicate device: a.
The samples from the clinical studies, in total 560 (275 from pMGN patients, 285 from control groups) were investigated for anti-PLA2R antibodies (IgG) using the two test systems EUROIMMUN Anti-PLA2R IFA and EUROIMMUN Anti-PLA2R ELISA (IgG). The discrepant samples were all from pMGN patients. Of the 25 discrepant samples positive with the EUROIMMUN Anti-PLA2R IFA and negative/borderline with the EUROIMMUN Anti-PLA2R ELISA (IgG), 19 samples exhibited a low IFA titer (1:10 to 1:32) and/or the ELISA result(s) was near cut-off (+/- 30%).

RU/mL:

		eonymon Aikiri Pack II A (LTCurren
		Positive	Borderline	Negative
EUROIMMUN	Positive	184			185
Anti-PLA2R ELISA	Borderline
(laG)	Negative	22		347	369
		212		348	560

EUROIMMIN Anti-DLA2D IEA (Prodic

Borderline defined as ≥14 to <20 RU/mL in the Instructions for Use. Borderline samples should be considered as potentially positive and retested.

Borderline samples counted as NEGATIVE:

		EUROIMMUN Anti-PLA2R IFA (Predicate)
		Positive	Negative
EUROIMMUNAnti-PLA2R ELISA(IgG)	Positive	184	1	185
	Negative	28	347	375
		212	348	560
Positive Agreement:	$184 / 212$	= 86.79%	95% C.I.:	81.5% - 91.0%
Negative Agreement:	$347 / 348$	= 99.71%	95% C.I.:	98.4% - 100.0%

Borderline samples counted as POSITIVE:

	EUROIMMUN Anti-PLA2R IFA (Predicate)
Positive	Negative

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EUROIMMUN®™™ US

EUROIMMUNAnti-PLA2R ELISA(IgG)	Positive	190	1	191
	Negative	22	347	369
		212	348	560
Positive Agreement:	$190 / 212$	= 89.62% 95% C.I.: 84.7% - 93.4%
Negative Agreement:	$347 / 348$	= 99.71% 95% C.I.: 98.4% - 100.0%

1 . . . . . . . .

።

... ..........................................................................................................................................................................

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Ratio:

		EUROIMMUN Anti-PLA2R IFA (Predicate)
		Positive	Borderline	Negative
EUROIMMUNAnti-PLA2R ELISA(IgG)	Positive	184	0	1	185
	Borderline	12	0	1	13
	Negative	16	0	346	362
		212	0	348	560

Borderline defined as ≥14 10 < 20 RUimLines for Use. Borderline samples should be considered as posentially positive and retested.

Borderline samples counted as NEGATIVE:

		EUROIMMUN Anti-PLA2R IFA (Predicate)
		Positive	Negative
EUROIMMUNAnti-PLA2R ELISA(IgG)	Positive	184	1	185
	Negative	28	347	375
		212	348	560
Positive Agreement:	184 / 212	= 86.79%	95% C.I.:	81.5% - 91.0%
Negative Agreement:	347 / 348	= 99.71%	95% C.I.:	98.4% - 100.0%

Borderline samples counted as POSITIVE:

		EUROIMMUN Anti-PLA2R IFA (Predicate)
		Positive	Negative
EUROIMMUNAnti-PLA2R ELISA(IgG)	Positive	196	2	198
	Negative	16	346	362
		212	348	560
Positive Agreement:	$196 / 212$	= 92.45%	95% C.I.: 88.0% - 95.6%
Negative Agreement:	$347 / 348$	= 99.71%	95% C.I.: 98.4% - 100.0%

b. Matrix comparison: Not Applicable

2. Clinical Study(s):

Clinical studies were performed in cooperation with different sites (see below). In total 560 clinically characterized samples (275 from pMGN patients, 285 from control groups) were investigated for anti-PLA2R antibodies (IgG). pMGN diagnosis was based on renal biopsy and was considered to be idiopathic/primary when no secondary cause of MN was suspected on the basis of clinical and laboratory criteria. The samples were drawn within 8 weeks after biopsy, before treatment; excluding patients who had been or were currently being treated with immunosuppressive drugs. With the EUROIMMUN Anti-PLA2R ELISA (IgG) using the 5-point calibrated analysis and a cut-off of 20 RU/ml, a sensitivity of 66.9% (95% C.I.: 61.0 - 72.4%) was found in pMGN, which is within the expected range of approximately 70% of anti-PLA2R as reported in the scientific literature. Specificity was 99.6% (95% C.I.: 98.1 - 100.0%).

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Sensitivity: a.

No. Panel			Anti-PLA2R ELISA (IgG)
			positive		95% C.I.
	l'rimary membranous glomerulonephritis(pMGN)	275	1845 borderline	66.9%	61.0-72.4%

Specificity: b.

No.	Panel	n	Anti-PLA2R ELISA (IgG)
			negative	%	95% C.I.
2	Secondary membranousglomerulonephritis (sMGN)	68	67	98.5%	92.1 - 100.0%
3	Non-membranous gromerulonephritides(GN)	63	63	100.0%	94.3 - 100.0%
4	Systemic lupus erythematosus (SLE)	30	30	100.0%	88.4 - 100.0%
5	Systemic sclerosis (SSc)	30	30	100.0%	88.4 - 100.0%
6	Psoriasis arthritis (PSA)	30	30	100.0%	88.4 - 100.0%
7	Rheumatoid arthritis (RA)	14	14	100.0%	76.8 - 100.0%
8	Thyreoiditis	50	50	100.0%	92.9 - 100.0%
	Total	285	284	99.6%	98.1 - 100.0%

Summary of Sensitivity & Specificity: C.

Clinical Samples(N = 560)	Clinical Diagnosis
	positive	negative	Total
EUROIMMUN Anti-PLA2RELISA (IgG)(RU/ml)	positive	184	1	185
	borderline	5	0	5
	negative	86	284	370
	Total	275	285	560

Borderline samples counted as negative:
Prevalence	$184 / 275 = 66.9%$	95% C.I.:	61.0% - 72.4%
Specificity	$284 / 285 = 99.6%$	95% C.I.:	98.1% - 100.0%
Borderline samples counted as positive:
Prevalence	$189 / 275 = 68.7%$	95% C.I.:	62.9% - 74.2%
Specificity	$284 / 285 = 99.6%$	95% C.I.:	98.1% - 100.0%

Clinical Samples(N = 560)	Clinical Diagnosis
	positive	negative	Total
EUROIMMUN Anti-PLA2RELISA (IgG)(Ratio)	positive	181	1	182
	borderline	9	1	10
	negative	85	283	288
	Total	275	285	560

Borderline samples counted as negative:
Prevalence	$181 / 275 = 65.8%$	95% C.I.:	59.9% - 71.4%
Specificity	$283 / 285 = 99.3%$	95% C.I.:	97.5% - 99.9%
Borderline samples counted as positive:
Prevalence	$190 / 275 = 69.1%$	95% C.I.:	63.3% - 74.5%
Specificity	$283 / 285 = 99.3%$	95% C.I.:	97.5% - 99.9%

Other clinical supportive data (when a. and b. are not applicable): d.

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1. Clinical Cut-off: See Assay Cut-off.
1. Expected Values/Reference Range:

European Donors: The levels of anti-PLA2R antibodies (IgG) were analyzed in a panel of 100 samples from apparently healthy blood donors (83 men and 17 women with an average age of 38 y; age range: 18 - 68 y).

Diagram & Table Expected Values/Reference Range (Europe)

Image /page/11/Figure/5 description: The image is a frequency distribution histogram. The x-axis represents RU/ml values ranging from 0.5 to 50, with specific values labeled such as 0.5, 0.8, 1.3, 2.0, 3.2, 5.0, 7.9, 13, 20, 32, and 50. The y-axis represents frequency, ranging from 0 to 30. The histogram shows the frequency of each RU/ml value, with the highest frequency occurring around 1.3 RU/ml.

n		100
positives		1
negatives			99
Prevalence			1.0 %
	RU/ml	Ratio
Lowest Value	0	0.0
Highest Value	32	1.6
Mean Value	2	0.1
Std Deviation	3.4	0.18

US Donors: The levels of anti-PLA2R antibodies (IgG) were analyzed in a panel of 248 samples from apparently healthy blood donors (151 men, 97 women, mean age 36 y, age range 17 - 50 y). The results are shown in the table below.

Diagram & Table Expected Values/Reference Range (USA) Frequency

Image /page/11/Figure/9 description: The image shows a frequency distribution histogram with the x-axis labeled in RU/ml from 0.5 to 50.0. The y-axis is labeled frequency from 0 to 60. The histogram shows the frequency of values, with the highest frequency occurring around 2.0 RU/ml. A table is also present with labels for n, positives, negatives, prevalence, lowest value, highest value, mean value, and standard deviation.

248

247

0.4 %

Ratio

0.0

1.6

0.1

2.8

RU/ml

2.8

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EUROIMMUN ાર

N. Proposed Labeling:

The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.

O. Conclusion:

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.

Printed NamelTitle

Michael Locke

Michael Locke/Dir. of Regulatory 6/27/14

Date

Signature

1100 The American Road · Morris Plains, NJ 07950 Tel: 973.656 1000/1.800.913.2022 : Fax: 973.656.1098 · E-mail: M.Locke@euroimmun.us

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Image /page/13/Picture/0 description: The image shows the logo for the Department of Health & Human Services USA. The logo is a circular seal with the words "DEPARTMENT OF HEALTH & HUMAN SERVICES USA" around the perimeter. Inside the circle is an image of an eagle with its wings spread. The eagle is facing to the right and has three wavy lines extending from its body.

DEPARTMENT OF HEALTH & HUMAN SERVICES

Public Health Service

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

June 27, 2014

EUROIMMUN US Inc. Michael Locke Director, Regulatory Affairs 1100 The American Road Morris Plains NJ 07950

Re: K132195

· Trade/Device Name; EUROIMMUN Anti-PLA2R ELISA (IgG) Regulation Number: 21 CFR 866.5780 Regulation Name: Anti-phospholipid A2 receptor immunological test system Regulatory Class: II Product Code: PGV Dated: June 9, 2014 Received: June 11, 2014

Dear Mr. Locke:

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 mislcading,

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 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device 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.

{14}------------------------------------------------

Page 2-Mr. Locke

If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education 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. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 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 Industry and Consumer Education 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,

Elizabeth A. Stafford -S

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

Enclosure

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Indications for Use

510(k) Number (if known) K132195

Device Name

EUROIMMUN Anti-PLA2R ELISA (IgG)

Indications for Use (Describe)

The EUROIMMUN Anti-PLA2R ELISA (IgG) test kit is intended for the qualitative determination of IgG class autoantibodies against phospholipase A2 receptor (PLA2R) in human serum. It is used as and in the diagnosis of primary membranous glomershriis (pMGN), in conjunction with other laboratory and clinical findings.

Type of Use (Select one or both, as applicable)

Prescription Use (Part 21 CFR 801 Subpart D)

_ Over-The-Counter Use (21 CFR 801 Subpart C)

PLEASE DO NOT WRITE BELOW THIS LINE - CONTINUE ON A SEPARATE PAGE IF NEEDED.

FOR FDA USE ONLY

Concurrence of Center for Devices and Radiological Health (CDRH) (Signature)

Elizabeth A. Stafford -S

This section applies only to requirements of the Paperwork Reduction Act of 1995.

*DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW."

The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:

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Form Approved: OMB No. 0910-0120 Expiration Date: January 31, 2017 See PRA Statement below.

Regulation Number and Section

§ 866.5780 Anti-phospholipase A2 receptor immunological test system.

(a)
Identification. An anti-phospholipase A2 receptor immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the autoantibodies in human blood samples that react with phospholipase A2 receptor. The measurements aid in the diagnosis of primary membranous glomerulonephritis (pMGN), in conjunction with other laboratory and clinical findings.(b)
Classification. Class II (special controls). The special controls for this device are:(1) Design verification and validation must include:
(i) A detailed description of the device that includes:
(A) A detailed description of all parts of the test system, including a description of the assay parts in the kit and all required ancillary reagents;
(B) A detailed description of instrumentation and equipment, and illustrations or photographs of non-standard equipment or methods if applicable;
(C) Detailed documentation of the device software, including standalone software applications and hardware-based devices that incorporate software where applicable;
(D) A detailed description of appropriate internal and external quality controls that are recommended or provided. The description must identify those control elements that are incorporated into the recommended testing procedures;
(E) Detailed specifications for sample collection, processing, and storage;
(F) A detailed description of methodology and assay procedure; and
(G) Detailed specification of the criteria for test results interpretation and reporting.
(ii) Information that demonstrates the performance characteristics of the device, including:
(A) Device precision/reproducibility data generated from within-run, between-run, between-day, between-lot, between-operator, between-instruments, between-site, and total precision for multiple nonconsecutive days as applicable. A well-characterized panel of patient samples or pools from the intended use population that covers the device measuring range must be used;
(B) Device linearity data generated from patient samples covering the assay measuring range if applicable;
(C) Information on traceability to a reference material and description of value assignment of calibrators and controls if applicable;
(D) Device analytical sensitivity data, including limit of blank, limit of detection, and limit of quantitation if applicable;
(E) Device analytical specificity data, including interference by endogenous and exogenous substances, as well as cross-reactivity with samples derived from patients with other autoimmune diseases or conditions;
(F) Device instrument carryover data when applicable;
(G) Device stability data including real-time stability under various storage times and temperatures;
(H) Specimen stability data including stability under various storage times, temperatures, freeze-thaw, and transport conditions where appropriate;
(I) Method comparison data generated by comparison of the results obtained with the device to those obtained with a legally marketed predicate device with similar indication of use. Patient samples from the intended use population covering the device measuring range must be used;
(J) Specimen matrix comparison data if more than one specimen type or anticoagulant can be tested with the device. Samples used for comparison must be from patient samples covering the device measuring range;
(K) A description of how the assay cut-off (the medical decision point between positive and negative) was established and validated as well as supporting data;
(L) A clinical performance assessment established by comparing data generated by testing samples from the intended use population and differential diagnosis groups with the device to the clinical diagnostic standard. Diagnosis of pMGN must be based primarily on clinical history, physical examination, laboratory tests (including urinalysis), and renal biopsy. Membranous glomerulonephritis is considered to be idiopathic/primary when no secondary cause can be elucidated on the basis of clinical and laboratory criteria. The differential diagnosis groups must include secondary membranous glomerulonephritis, membranoproliferative glomerulonephritis, lupus nephritis, focal segmental glomerulosclerosis, IgA nephritis, diabetic nephropathy, systemic lupus erythematosus, systemic sclerosis, and Goodpasture syndrome. Diagnosis of autoimmune and immune-mediated diseases that are associated with membranous glomerulonephritis must be based on established diagnostic criteria and clinical evaluation. For all samples, clinical criteria, including demographic information, must be considered in the differentiation between pMGN and secondary membranous glomerulonephritis. The clinical validation results must demonstrate correlation clinical sensitivity and clinical specificity between the test values and the presence or absence of pMGN. The data must be summarized in tabular format comparing the interpretation of results to the disease status; and
(M) Expected/reference values generated by testing an adequate number of samples from apparently healthy normal individuals.
(iii) Identification of risk mitigation elements used by the device, including a description of all additional procedures, methods, and practices incorporated into the directions for use that mitigate risks associated with testing.
(2) The label required under § 809.10(a) and labeling required under § 809.10(b) of this chapter must include warnings relevant to the assay including:
(i) A warning statement that explains: The device is for use by laboratory professionals in a clinical laboratory setting;
(ii) A warning statement that explains: The test is not a standalone test but an adjunct to other clinical information. A diagnosis of pMGN or secondary MGN should not be made based on a single test result. The clinical symptoms, results on physical examination, and laboratory tests (
e.g., serological tests), when appropriate, should always be taken into account when considering the diagnosis of primary versus secondary MGN;(iii) A warning statement that explains: Absence of circulating PLA2R autoantibody does not rule out a diagnosis of pMGN; and
(iv) A warning statement that explains: The assay has not been demonstrated to be effective for monitoring the stage of disease or its response to treatment.
(3) The labeling required under § 809.10(b) of this chapter must include a description of the protocol and performance studies performed in accordance with paragraph (b)(1)(ii) of this section and a summary of the results.