The EUROIMMUN Anti-nRNP/Sm ELISA (IgG) test kit is intended for the qualitative determination of IgG class autoantibodies against nRNP/Sm in human serum and plasma (EDTA, Li-heparin, Citrate). It is used as an aid in the diagnosis of mixed connective tissue diseases and systemic lupus erythematosus, in conjunction with other laboratory and clinical findings.

The EUROIMMUN Anti-Sm ELISA (IgG) test kit is intended for the qualitative of IgG class autoantibodies against Sm in human serum and plasma (EDTA, Li-heparin, Citrate). It is used as an aid in the diagnosis of systemic lupus erythematosus, in conjunction with other laboratory and clinical findings.

The EUROIMMUN Anti-SS-A ELISA (IgG) test kit is intended for the qualitative determination of IgG class autoantibodies against SS-A in human serum and plasma (EDTA, Li-heparin, Citrate). It is used as an aid in the diagnosis of Sjögren's syndrome and systemic lupus erythematosus, in conjunction with other laboratory and clinical findings.

The EUROIMMUN Anti-SS-B ELISA (IgG) test kit is intended for the qualitative determination of IgG class autoantibodies against SS-B in human serum and plasma (EDTA, Li-heparin, Citrate). It is used as an aid in the diagnosis of Sjögren's syndrome and systemic lupus erythematosus, in conjunction with other laboratory and clinical findings.

The EUROIMMUN Anti-Scl-70 ELISA (IgG) test kit is intended for the qualitative determination of IgG class autoantibodies against Scl-70 in human serum and plasma (EDTA, Li-heparin, Citrate), It is used as an aid in the diagnosis of progressive systemic sclerosis, in conjunction with other laboratory and clinical findings.

The EUROIMMUN Anti-Centromeres ELISA (IgG) test kit is intended for the qualitative determination of IgG class autoantibodies against Centromeres in human serum and plasma (EDTA, Li-heparin, Citrate). It is used as an aid in the diagnosis of limited form of progressive systemic sclerosis (CREST syndrome), in conjunction with other laboratory and clinical findings.

The EUROIMMUN Anti-Jo-1 ELISA (IgG) test kit is intended for the qualitative determination of IgG class autoantibodies against Jo-1 in human serum and plasma (EDTA, Li-heparin, Citrate). It is used as an aid in the diagnosis of polymyositis and dermatomyositis, in conjunction with other laboratory and clinical findings.

The EUROIMMUN Anti-ribosomal P-proteins ELISA (IgG) test kit is intended for the qualitative determination of IgG class autoantibodies against ribosomal P-proteins in human serum and plasma (EDTA, Li-heparin, Citrate). It is used as an aid in the diagnosis of systemic lupus erythematosus, in conjunction with other laboratory and clinical findings.

Device Description

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

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

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

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

The EUROIMMUN Anti-Scl-70 ELISA (IgG) consists of a microwell ELISA plate coated with Scl-70 antigen, calibrator, positive and negative control, peroxidase-labelled anti-human IgG conjugate, sample buffer, wash buffer concentrate, TMB chromogen/substrate solution and stop solution.

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

The EUROIMMUN Anti-Jo-1 ELISA (IgG) consists of a microwell ELISA plate coated with Jo-1 antigen, calibrator, positive and negative control, peroxidase-labelled anti-human IgG conjugate, sample buffer, wash buffer concentrate, TMB chromogen/substrate solution and stop solution.

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

AI/ML Overview

Here's a breakdown of the acceptance criteria and study information for each of the EUROIMMUN ELISA kits for autoantibodies, as presented in the provided document.

It's important to note that the document outlines several ELISA kits (Anti-nRNP/Sm, Anti-Sm, Anti-SS-A, Anti-SS-B, Anti-Scl-70, Anti-Centromeres, and Anti-Jo-1, Anti-ribosomal P-proteins). Each kit has its own performance characteristics, and the requested information will be presented for each.

General Acceptance Criteria & Study Information (Applicable to all kits)

Type of Test: Qualitative enzyme immunoassay (ELISA) for IgG class autoantibodies.
Assay Cut-off: Ratio 1.0 (for all EUROIMMUN ELISA kits).
Sample Types: Human serum and plasma (EDTA, Li-heparin, Citrate).
Calibration: Relative evaluation/units.
Ground Truth for Training/Testing: Clinically characterized samples. The document does not explicitly separate training and test sets with distinct ground truth methods for each. Instead, "comparison studies" utilize a test set of clinically characterized samples against a predicate device, and "clinical studies" evaluate performance against clinical diagnosis.
Number of Experts to Establish Ground Truth: Not specifically mentioned for clinical characterization, but it's implied that clinical findings and diagnoses are established by medical professionals.
Qualifications of Experts: Not explicitly stated, but assumed to be relevant medical specialists for the diagnoses of the conditions studied (e.g., rheumatologists, immunologists).
Adjudication Method: Not explicitly mentioned. Clinical diagnoses are typically consensus-based among healthcare professionals or based on established diagnostic criteria.
MRMC Comparative Effectiveness Study: No MRMC studies were performed in this context. The comparisons are between the new ELISA device and a predicate ELISA device (algorithm only, as they are immunoassay kits) and against clinical diagnoses (standalone performance).
Standalone Performance: Yes, the clinical studies evaluate the standalone performance of the algorithm (the ELISA kit) in terms of clinical sensitivity and specificity against clinical diagnoses.
Data Provenance: Samples were obtained from different sources from Europe and North America (retrospective, as they are "clinically characterized samples").
Sample Size for Training Set: Not explicitly stated. The document focuses on performance characteristics and comparison/clinical studies with given sample sizes. It is common for ELISA development to use a subset of samples for initial optimization and cut-off determination, but specific "training set" sizes are not detailed in this regulatory summary.

1. EUROIMMUN Anti-nRNP/Sm ELISA (IgG)

Intended Use: Qualitative determination of IgG class autoantibodies against nRNP/Sm in human serum and plasma (EDTA, Li-heparin, Citrate) as an aid in diagnosing mixed connective tissue diseases and systemic lupus erythematosus.

Table of Acceptance Criteria and Reported Device Performance:

Acceptance Criteria (Implied)	Reported Device Performance
High correlation with Predicate Device	Method Comparison (vs. Inova Quanta Lite RNP ELISA):Overall agreement: 97.2% (95% C.I.: 94.6% - 98.8%)Positive agreement: 91.2% (95% C.I.: 83.4% - 96.1%)Negative agreement: 100.0% (95% C.I.: 98.1% - 100.0%)
Acceptable Clinical Sensitivity in target diseases	Clinical Sensitivity:Mixed connective tissue diseases (MCTD): 100.0% (95% C.I.: 94.5% - 100.0%) (n=65)Systemic lupus erythematosus (SLE): 23.3% (95% C.I.: 19.2% - 27.7%) (n=404)
Acceptable Clinical Specificity in control groups	Clinical Specificity (Total control groups, excluding myositis):99.3% (95% C.I.: 98.0% - 99.9%) (Total n=577, excluding myositis)Individual specificities: Rheumatoid arthritis (100.0%), Systemic sclerosis (100.0%), Sjögren's syndrome (97.7%), Other autoimmune diseases (98.4%), Borreliosis (100.0%)
Good analytical precision (Intra-assay and Inter-assay)	Intra-assay reproducibility: 100% agreement for positive/negative samples with various ratios (n=20 per sample)Inter-assay reproducibility: 100% agreement for 5/6 samples, one negative sample 97.5% negative (n=40 per sample)Lot-to-lot reproducibility: High agreement reported, with expected classification (positive/negative) maintained across lots.
No significant cross-reactivity with other autoantibodies	Cross-reactivity: All 30 sera positive for various other autoantibodies (rib.P-P, SS-A, SS-B, Scl-70, Jo-1, centromeres, Rubella, Measles, HSV1, Borrelia) were negative in Anti-nRNP/Sm ELISA.
No significant interference from common interfering substances	Interference: Individual recovery within 85-109% for hemoglobin (up to 1000 mg/dl), triglyceride (up to 2000 mg/dl), bilirubin (up to 40 mg/dl), and rheumatoid factor (up to 500 IU/ml).
Equivalence across different plasma types vs. serum	Matrix comparison: Coefficients of determination (R²) >0.99, Mean %recovery 101-105%, Range of %recovery 91-120% (for EDTA, Li-heparin, Citrate plasma vs. serum, n=15 pairs each). Regression equations near ideal (intercept 0, slope 1).

2. EUROIMMUN Anti-Sm ELISA (IgG)

Intended Use: Qualitative determination of IgG class autoantibodies against Sm in human serum and plasma (EDTA, Li-heparin, Citrate) as an aid in diagnosing systemic lupus erythematosus.

Table of Acceptance Criteria and Reported Device Performance:

Acceptance Criteria (Implied)	Reported Device Performance
High correlation with Predicate Device	Method Comparison (vs. Inova Quanta Lite Sm ELISA):Overall agreement: 95.9% (95% C.I.: 93.0% - 97.9%)Positive agreement: 84.1% (95% C.I.: 69.9% - 93.4%)Negative agreement: 98.0% (95% C.I.: 95.4% - 99.3%)
Acceptable Clinical Sensitivity in target diseases	Clinical Sensitivity:Systemic lupus erythematosus (SLE): 11.4% (95% C.I.: 8.5% - 14.8%) (n=414)
Acceptable Clinical Specificity in control groups	Clinical Specificity (Total control groups):99.0% (95% C.I.: 97.9% - 99.6%) (Total n=622)Individual specificities: Rheumatoid arthritis (100.0%), Systemic sclerosis (100.0%), Sjögren's syndrome (100.0%), Polymyositis/dermatomyositis (100.0%), Mixed connective tissue diseases (86.7%), Other autoimmune diseases (100.0%), Borreliosis (100.0%)
Good analytical precision (Intra-assay and Inter-assay)	Intra-assay reproducibility: 100% agreement for positive/negative samples (n=20 per sample)Inter-assay reproducibility: 100% agreement for 4/6 samples, one negative 97.5% negative, another 0% negative (n=40 per sample)Lot-to-lot reproducibility: High agreement reported, with expected classification (positive/negative) maintained across lots.
No significant cross-reactivity with other autoantibodies	Cross-reactivity: All 30 sera positive for various other autoantibodies (rib.P-P, nRNP/Sm, SS-A, Scl-70, Jo-1, centromeres, Rubella, Measles, HSV1, Borrelia) were negative in Anti-Sm ELISA.
No significant interference from common interfering substances	Interference: Individual recovery within 90-111% for hemoglobin (up to 1000 mg/dl), triglyceride (up to 2000 mg/dl), bilirubin (up to 40 mg/dl), and rheumatoid factor (up to 500 IU/ml).
Equivalence across different plasma types vs. serum	Matrix comparison: Coefficients of determination (R²) >0.99, Mean %recovery 99-103%, Range of %recovery 89-118% (for EDTA, Li-heparin, Citrate plasma vs. serum, n=16 pairs each). Regression equations near ideal (intercept 0, slope 1).

3. EUROIMMUN Anti-SS-A ELISA (IgG)

Intended Use: Qualitative determination of IgG class autoantibodies against SS-A in human serum and plasma (EDTA, Li-heparin, Citrate) as an aid in diagnosing Sjögren's syndrome and systemic lupus erythematosus.

Table of Acceptance Criteria and Reported Device Performance:

Acceptance Criteria (Implied)	Reported Device Performance
High correlation with Predicate Device	Method Comparison (vs. Inova Quanta Lite SS-A ELISA):Overall agreement: 96.1% (95% C.I.: 93.2% - 98.0%)Positive agreement: 92.8% (95% C.I.: 86.8% - 96.7%)Negative agreement: 98.3% (95% C.I.: 95.2% - 99.7%)
Acceptable Clinical Sensitivity in target diseases	Clinical Sensitivity:Sjögren's syndrome: 73.9% (95% C.I.: 63.4% - 82.7%) (n=88)Systemic lupus erythematosus (SLE): 40.6% (95% C.I.: 35.8% - 45.6%) (n=404)
Acceptable Clinical Specificity in control groups	Clinical Specificity (Total control groups):94.8% (95% C.I.: 92.5% - 96.5%) (Total n=534)Individual specificities: Systemic sclerosis (92.6%), Polymyositis/dermatomyositis (91.4%), Rheumatoid arthritis (97.0%), Mixed connective tissue diseases (91.1%), Other autoimmune diseases (100.0%), Borreliosis (100.0%)
Good analytical precision (Intra-assay and Inter-assay)	Intra-assay reproducibility: 100% agreement for positive/negative samples (n=20 per sample)Inter-assay reproducibility: 100% agreement for positive/negative samples (n=40 per sample)Lot-to-lot reproducibility: High agreement reported, with expected classification (positive/negative) maintained across lots.
No significant cross-reactivity with other autoantibodies	Cross-reactivity: All 30 sera positive for various other autoantibodies (rib.P-P, nRNP/Sm, Sm, Scl-70, Jo-1, centromeres, Rubella, Measles, HSV1, Borrelia) were negative in Anti-SS-A ELISA.
No significant interference from common interfering substances	Interference: Individual recovery within 93-107% for hemoglobin (up to 1000 mg/dl), triglyceride (up to 2000 mg/dl), bilirubin (up to 40 mg/dl), and rheumatoid factor (up to 500 IU/ml).
Equivalence across different plasma types vs. serum	Matrix comparison: Coefficients of determination (R²) >0.99, Mean %recovery 95-98%, Range of %recovery 85-104% (for EDTA, Li-heparin, Citrate plasma vs. serum, n=16 pairs each). Regression equations near ideal (intercept 0, slope 1).

4. EUROIMMUN Anti-SS-B ELISA (IgG)

Intended Use: Qualitative determination of IgG class autoantibodies against SS-B in human serum and plasma (EDTA, Li-heparin, Citrate) as an aid in diagnosing Sjögren's syndrome and systemic lupus erythematosus.

Table of Acceptance Criteria and Reported Device Performance:

Acceptance Criteria (Implied)	Reported Device Performance
High correlation with Predicate Device	Method Comparison (vs. Inova Quanta Lite SS-B ELISA):Overall agreement: 98.5% (95% C.I.: 96.3% - 99.6%)Positive agreement: 96.5% (95% C.I.: 90.0% - 99.3%)Negative agreement: 99.5% (95% C.I.: 97.1% - 100.0%)
Acceptable Clinical Sensitivity in target diseases	Clinical Sensitivity:Sjögren's syndrome: 39.8% (95% C.I.: 29.5% - 50.8%) (n=88)Systemic lupus erythematosus (SLE): 13.9% (95% C.I.: 10.6% - 17.6%) (n=404)
Acceptable Clinical Specificity in control groups	Clinical Specificity (Total control groups):98.1% (95% C.I.: 96.6% - 99.1%) (Total n=534)Individual specificities: Rheumatoid arthritis (99.4%), Systemic sclerosis (95.1%), Polymyositis/dermatomyositis (98.7%), Mixed connective tissue diseases (93.3%), Other autoimmune diseases (100.0%), Borreliosis (100.0%)
Good analytical precision (Intra-assay and Inter-assay)	Intra-assay reproducibility: 100% agreement for positive/negative samples (n=20 per sample)Inter-assay reproducibility: 100% agreement for positive/negative samples (n=40 per sample)Lot-to-lot reproducibility: High agreement reported, with expected classification (positive/negative) maintained across lots.
No significant cross-reactivity with other autoantibodies	Cross-reactivity: All 30 sera positive for various other autoantibodies (rib.P-P, nRNP/Sm, Sm, SS-A, Scl-70, Jo-1, centromeres, Rubella, Measles, HSV1, Borrelia) were negative in Anti-SS-B ELISA.
No significant interference from common interfering substances	Interference: Individual recovery within 92-116% for hemoglobin (up to 1000 mg/dl), triglyceride (up to 2000 mg/dl), bilirubin (up to 40 mg/dl), and rheumatoid factor (up to 500 IU/ml).
Equivalence across different plasma types vs. serum	Matrix comparison: Coefficients of determination (R²) >0.99, Mean %recovery 100-105%, Range of %recovery 78-116% (for EDTA, Li-heparin, Citrate plasma vs. serum, n=16 pairs each). Regression equations near ideal (intercept 0, slope 1).

5. EUROIMMUN Anti-Scl-70 ELISA (IgG)

Intended Use: Qualitative determination of IgG class autoantibodies against Scl-70 in human serum and plasma (EDTA, Li-heparin, Citrate) as an aid in diagnosing progressive systemic sclerosis.

Table of Acceptance Criteria and Reported Device Performance:

Acceptance Criteria (Implied)	Reported Device Performance
High correlation with Predicate Device	Method Comparison (vs. Inova Quanta Lite Scl-70 ELISA):Overall agreement: 100.0% (95% C.I.: 98.8% - 100.0%)Positive agreement: 100.0% (95% C.I.: 97.1% - 100.0%)Negative agreement: 100.0% (95% C.I.: 98.0% - 100.0%)
Acceptable Clinical Sensitivity in target diseases	Clinical Sensitivity:Systemic sclerosis: 23.2% (95% C.I.: 18.4% - 28.6%) (n=280)Diffuse systemic sclerosis: 59.4% (95% C.I.: 48.9% - 69.3%) (n=96)Limited systemic sclerosis: 5.3% (95% C.I.: 2.0% - 11.2%) (n=113)
Acceptable Clinical Specificity in control groups	Clinical Specificity (Total control groups):99.8% (95% C.I.: 99.1% - 100.0%) (Total n=629)Individual specificities: Systemic lupus erythematosus (100.0%), Polymyositis/dermatomyositis (100.0%), Rheumatoid arthritis (99.4%), Sjögren's syndrome (100.0%), Mixed connective tissue diseases (100.0%), Other autoimmune diseases (100.0%), Borreliosis (100.0%)
Good analytical precision (Intra-assay and Inter-assay)	Intra-assay reproducibility: 100% agreement for positive/negative samples (n=20 per sample)Inter-assay reproducibility: 100% agreement for positive/negative samples (n=40 per sample)Lot-to-lot reproducibility: High agreement reported, with expected classification (positive/negative) maintained across lots.
No significant cross-reactivity with other autoantibodies	Cross-reactivity: All 30 sera positive for various other autoantibodies (rib.P-P, nRNP/Sm, Sm, SS-A, SS-B, Jo-1, centromeres, Rubella, Measles, HSV1, Borrelia) were negative in Anti-Scl-70 ELISA.
No significant interference from common interfering substances	Interference: Individual recovery within 86-109% for hemoglobin (up to 1000 mg/dl), triglyceride (up to 2000 mg/dl), bilirubin (up to 40 mg/dl), and rheumatoid factor (up to 500 IU/ml).
Equivalence across different plasma types vs. serum	Matrix comparison: Coefficients of determination (R²) >0.98, Mean %recovery 96-101%, Range of %recovery 86-113% (for EDTA, Li-heparin, Citrate plasma vs. serum, n=16 pairs each). Regression equations near ideal (intercept 0, slope 1).

6. EUROIMMUN Anti-Centromeres ELISA (IgG)

Intended Use: Qualitative determination of IgG class autoantibodies against Centromeres in human serum and plasma (EDTA, Li-heparin, Citrate) as an aid in diagnosing limited form of progressive systemic sclerosis (CREST syndrome).

Table of Acceptance Criteria and Reported Device Performance:

Acceptance Criteria (Implied)	Reported Device Performance
High correlation with Predicate Device	Method Comparison (vs. Inova Quanta Lite Centromeres ELISA):Overall agreement: 98.3% (95% C.I.: 96.1% - 99.5%)Positive agreement: 93.6% (95% C.I.: 85.7% - 97.9%)Negative agreement: 100.0% (95% C.I.: 98.3% - 100.0%)
Acceptable Clinical Sensitivity in target diseases	Clinical Sensitivity:Systemic sclerosis: 37.5% (95% C.I.: 31.8% - 43.5%) (n=280)Diffuse systemic sclerosis: 7.3% (95% C.I.: 3.0% - 14.4%) (n=96)Limited systemic sclerosis: 74.3% (95% C.I.: 65.3% - 82.1%) (n=113)
Acceptable Clinical Specificity in control groups	Clinical Specificity (Total control groups):99.0% (95% C.I.: 97.8% - 99.6%) (Total n=597)Individual specificities: Systemic lupus erythematosus (99.4%), Polymyositis/dermatomyositis (100.0%), Rheumatoid arthritis (99.4%), Sjögren's syndrome (96.6%), Mixed connective tissue diseases (97.8%), Other autoimmune diseases (100.0%), Borreliosis (100.0%)
Good analytical precision (Intra-assay and Inter-assay)	Intra-assay reproducibility: 100% agreement for positive/negative samples (n=20 per sample)Inter-assay reproducibility: 100% agreement for positive/negative samples (n=40 per sample)Lot-to-lot reproducibility: High agreement reported, with expected classification (positive/negative) maintained across lots.
No significant cross-reactivity with other autoantibodies	Cross-reactivity: All 30 sera positive for various other autoantibodies (rib.P-P, nRNP/Sm, Sm, SS-A, Scl-70, Jo-1, Rubella, Measles, HSV1, Borrelia) were negative in Anti-Centromeres ELISA.
No significant interference from common interfering substances	Interference: Individual recovery within 91-111% for hemoglobin (up to 1000 mg/dl), triglyceride (up to 2000 mg/dl), bilirubin (up to 40 mg/dl), and rheumatoid factor (up to 500 IU/ml).
Equivalence across different plasma types vs. serum	Matrix comparison: Coefficients of determination (R²) >0.99, Mean %recovery 93-98%, Range of %recovery 83-108% (for EDTA, Li-heparin, Citrate plasma vs. serum, n=16 pairs each). Regression equations near ideal (intercept 0, slope 1).

7. EUROIMMUN Anti-Jo-1 ELISA (IgG)

Intended Use: Qualitative determination of IgG class autoantibodies against Jo-1 in human serum and plasma (EDTA, Li-heparin, Citrate) as an aid in diagnosing polymyositis and dermatomyositis.

Table of Acceptance Criteria and Reported Device Performance:

Acceptance Criteria (Implied)	Reported Device Performance
High correlation with Predicate Device	Method Comparison (vs. Inova Quanta Lite Jo-1 ELISA):Overall agreement: 99.3% (95% C.I.: 97.6% - 99.9%)Positive agreement: 98.5% (95% C.I.: 91.7% - 100.0%)Negative agreement: 99.6% (95% C.I.: 97.6% - 100.0%)
Acceptable Clinical Sensitivity in target diseases	Clinical Sensitivity:Myositis (Polymyositis/dermatomyositis): 18.6% (95% C.I.: 13.2% - 25.2%) (n=177)
Acceptable Clinical Specificity in control groups	Clinical Specificity (Total control groups):99.6% (95% C.I.: 98.8% - 99.9%) (Total n=699)Individual specificities: Systemic lupus erythematosus (99.1%), Rheumatoid arthritis (99.4%), Systemic sclerosis (100.0%), Sjögren's syndrome (100.0%), Mixed connective tissue diseases (100.0%), Fibromyalgia (100.0%), Other autoimmune diseases (100.0%), Borreliosis (100.0%)
Good analytical precision (Intra-assay and Inter-assay)	Intra-assay reproducibility: 100% agreement for positive/negative samples (n=20 per sample)Inter-assay reproducibility: 100% agreement for positive/negative samples (n=40 per sample)Lot-to-lot reproducibility: High agreement reported, with expected classification (positive/negative) maintained across lots.
No significant cross-reactivity with other autoantibodies	Cross-reactivity: All 30 sera positive for various other autoantibodies (rib.P-P, nRNP/Sm, Sm, SS-A, SS-B, Scl-70, centromeres, Rubella, Measles, HSV1, Borrelia) were negative in Anti-Jo-1 ELISA.
No significant interference from common interfering substances	Interference: Individual recovery within 91-122% for hemoglobin (up to 1000 mg/dl), triglyceride (up to 2000 mg/dl), bilirubin (up to 40 mg/dl), and rheumatoid factor (up to 500 IU/ml).
Equivalence across different plasma types vs. serum	Matrix comparison: Coefficients of determination (R²) >0.98, Mean %recovery 95-103%, Range of %recovery 85-117% (for EDTA, Li-heparin, Citrate plasma vs. serum, n=15 pairs each). Regression equations near ideal (intercept 0, slope 1).

8. EUROIMMUN Anti-ribosomal P-proteins ELISA (IgG)

Intended Use: Qualitative determination of IgG class autoantibodies against ribosomal P-proteins in human serum and plasma (EDTA, Li-heparin, Citrate) as an aid in diagnosing systemic lupus erythematosus.

Table of Acceptance Criteria and Reported Device Performance:

Acceptance Criteria (Implied)	Reported Device Performance
High correlation with Predicate Device	Method Comparison (vs. Inova Quanta Lite Ribosomal P ELISA):Overall agreement: 95.9% (95% C.I.: 92.6% - 98.0%)Positive agreement: 100.0% (95% C.I.: 87.7% - 100.0%)Negative agreement: 95.3% (95% C.I.: 91.6% - 97.7%)
Acceptable Clinical Sensitivity in target diseases	Clinical Sensitivity:Systemic lupus erythematosus (SLE): 5.3% (95% C.I.: 3.3% - 8.1%) (n=376)
Acceptable Clinical Specificity in control groups	Clinical Specificity (Total control groups):99.2% (95% C.I.: 98.0% - 99.8%) (Total n=500)Individual specificities: Polymyositis/dermatomyositis (98.7%), Rheumatoid arthritis (100.0%), Systemic sclerosis (100.0%), Sjögren's syndrome (98.2%), Mixed connective tissue diseases (97.8%), Other autoimmune diseases (100.0%), Borreliosis (100.0%)
Good analytical precision (Intra-assay and Inter-assay)	Intra-assay reproducibility: 100% agreement for positive/negative samples (n=20 per sample)Inter-assay reproducibility: 100% agreement for 5/6 samples, one negative 97.5% negative (n=40 per sample)Lot-to-lot reproducibility: High agreement reported, with expected classification (positive/negative) maintained across lots.
No significant cross-reactivity with other autoantibodies	Cross-reactivity: All 30 sera positive for various other autoantibodies (nRNP/Sm, Sm, SS-A, Scl-70, Jo-1, centromeres, Rubella, Measles, HSV1, Borrelia) were negative in Anti-ribosomal P-proteins ELISA.
No significant interference from common interfering substances	Interference: Individual recovery within 84-115% for hemoglobin (up to 1000 mg/dl), triglyceride (up to 2000 mg/dl), bilirubin (up to 40 mg/dl), and rheumatoid factor (up to 500 IU/ml).
Equivalence across different plasma types vs. serum	Matrix comparison: Coefficients of determination (R²) >0.99, Mean %recovery 95-98%, Range of %recovery 79-107% (for EDTA, Li-heparin, Citrate plasma vs. serum, n=16 pairs each). Regression equations near ideal (intercept 0, slope 1).

Summary

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K 1.23261

JUN 1 2 2013

ATTACHMENT I

PREMARKET NOTIFICATION 210(K) SAFETY AND EFFECTIVENESS SUMMARY (as required by 21 CFR § 807.92)

A. 510(k)Number: K123261
B. Purpose for Submission: New device
C. Measurand:

Anti-nRNP/Sm autoantibodies
Type of Test: Qualitative enzyme immunoassay
E. Applicant:
- EUROIMMUN US INC.
Proprietary and Established Names: ಿ. EUROIMMUN Anti-nRNP/Sm ELISA (IgG)
G. Requlatory Information:
- Regulation: 1.
  - 21 CFR 866.5110 Antinuclear antibody immunological test system
- 1. Classification: Class II
- 1. Product code: LKO
- 1. Panel:

Immunology Intended Use:

Intended use(s): 1 .
The EUROIMMUN Anti-nRNP/Sm ELISA (IgG) test kit is intended for the qualitative determination of IgG class autoantibodies against nRNP/Sm in human serum and plasma (EDTA, Li-heparin, Citrate). It is used as an aid in the diagnosis of mixed connective tissue diseases and systemic lupus erythematosus, in conjunction with other laboratory and clinical findings.
1. Indication(s) for use: 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.

l. Device Description:

J. Substantial Equivalence Information:

Predicate device name (s): 1 .
- Inova Quanta Lite RNP ELISA
1. Predicate 510(k) number(s): K922833

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3. Comparison with predicate:

Item	New Device	Predicate Device
Intended use	Detection of IgG antibodies to nRNP/Sm	Same
Technology	ELISA	Same
Assay platform	96-well microtiter plates	Same
Calibration	Relative evaluation	Same
Antigen	Purified U1-nRNP complex; U1-nRNP contains RNP aswell as Sm reactive proteins	Same
Conjugate	Anti-human IgG labeled with horseradish peroxidase	Same
Substrate	TMB	Same
Reagent preparation	All reagents, calibrators and controls are ready to use,except for the wash buffer.	Same
Procedure	Sample incubation with micro-well antigen coated plate,followed by a wash step, incubation with an anti-humanIgG enzyme conjugate; wash step, incubation withsubstrate; then the addition of a stop solution andreading at 450nm.	Same
Item	New Device	Predicate Device
Assay format	Qualitative	"Semi-quantitative" (using low positive control)
Sample dilution	1:201	1:101
Calibrators andcontrols	1 calibrator2 controls: 1 positive, 1 negative	3 controls: 1 high positive, 1 low positive (used forcalculation of results), 1 negative
Wash buffer	10x concentrate	40x concentrate
Stop solution	0.5 M sulphuric acid	0.344 M sulphuric acid
Sample types	Serum or plasma (EDTA, Li-heparin, Citrate)	Serum
Reported results	Ratio	Units
Cut off level	Ratio 1.0	20 Units

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

Guidance for Industry and FDA Staff: Recommendations for Antibody (ANA) Test System Premarket (510(k)) Submissions (January 22, 2009)

L. Test Principle:

Patient samples are diluted 1:201 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 µl 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 pl of wash buffer to remove any unbound enzyme conjugate and 100 µl of the chromogen substrate is added. The strips are incubated for 15 minutes at room temperature and 100 µl 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.

Performance Characteristics (where applicable): M.

Analytical performance:
- Precision/Reproducibility: a.

The reproducibility of the test was investigated using sera with different concentrations. Intra-assay reproducibility is based on 20 determinations and inter-assay reproducibility on 40 determinations performed in 10 different runs on 5 days with 2 runs per day according to the package insert. The following results were obtained:

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Intra-assay reproducibility

n = 20	Anti-nRNP/Sm ELISA (IgG) Ratio
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6
Mean value (x):	5.1	3.3	1.9	1.2	0.8	0.4
Range of values:	5.0 - 5.1	3.2 - 3.4	1.8 - 2.0	1.1 - 1.2	0.7 - 0.9	0.3 - 0.4
Expected result:	positive	positive	positive	positive	negative	negative
% positive:	100%	100%	100%	100%	0%	0%
% negative:	0%	0%	0%	0%	100%	100%

Inter-assay reproducibility

	Anti-nRNP/Sm ELISA (IgG)
n = 10 x 4	Ratio
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6
Mean value (x):	5.2	3.3	1.8	1.1	0.8	0.4
Range of values:	4.8 - 5.5	3.1 - 3.6	1.6 - 1.9	1.1 - 1.2	0.7 - 0.9	0.3 - 0.4
Expected result:	positive	positive	positive	positive	negative	negative
% positive:	100%	100%	100%	100%	0%	0%
% negative:	0%	0%	0%	0%	100%	100%

The lot to lot reproducibility was investigated during the validaton and quality control of the kit using different lots with QC samples distributed over the measurement range. The following results were obtained:

Lot to lot reproducibility

	Anti-nRNP/Sm ELISA (IgG)Ratio							Anti-nRNP/Sm ELISA (IgG)Ratio
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6		Sample 7	Sample 8	Sample 9	Sample 10	Sample 11
n	6*	6*	6*	6*	6*	6*	n	11**	11**	11**	11**	11**
Mean value (x):	3.6	2.5	5.4	0.4	0.9	1.1	Mean value (x):	0.2	3.4	5.2	6.6	8.5
Range of values:	3.4 - 3.9	2.3 – 2.8	4.8 - 5.7	0.3 - 0.4	0.9 - 0.9	1.1 - 1.1	Range of values:	0.1 - 0.3	3.1 - 3.8	4.7 - 5.9	5.7 - 7.2	8.0 - 9.7
Expected result:	positive	positive	positive	negative	negative	positive	Expected result:	negative	positive	positive	positive	positive
% positive:	100%	100%	100%	0%	0%	100%	% positive:	0%	100%	100%	100%	100%
% negative:	0%	0%	0%	100%	100%	0%	% negative:	100%	0%	0%	0%	0%

*3 lots x 2 runs ** n lots x 1 run

b. Linearity/assay reportable range: Not applicable.

Not applicable.
High dose/leak-off.

C. High dose Hook effect

Potential for a high dose Hook effect is a phenomenon that is inherent with one step "sandwich" assay designs: Very high concentrations of antigen in the patient sample bind to all available sites saturating them - on both the antibody-solid phase and the antibody-labeled conjugate and thereby prevent the "sandwich" formation. Under these conditions, the measured level of analyte may be significantly lower than the actual level present in the sample. The two-step immunoassay design of the Anti-nRNP/Sm ELISA (IgG) eliminates the adverse contribution of binding proteins, endogenous interfering substances and general matrix effects due to the extra wash step.

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Image /page/3/Picture/1 description: The image shows a pattern of alternating black and white shapes. The black shapes are arranged in a row and are separated by white spaces. The black shapes are rounded and symmetrical, resembling beads or capsules. The white spaces between the black shapes are rectangular and uniform in size.

d. Traceability, Stability, Expected values (controls, calibrators or methods):

A recognized standard or reference material for anti-nuclear antibodies is not available. Results of this assay are given in ratios. The reactivity of the Anti-nRNP/Sm ELISA was verified using the ANA reference panel of the CDC Centers for Disease Control and Prevention, Atlanta, USA.

Limit of detection: e.
Not applicable.
f Analytical specificity:

Cross-reactivity: The quality of the antigen coated on the plates ensures a high specificity of the ELISA. Cross reactivity was investigated using a panel of 30 sera serologically positive for antibodies against rib.P-P, SS-A, SS-B, Scl-70, Jo-1, centromeres, Rubella virus, Measles virus, Herpes simplex virus type 1 and Borrelia burgdorferi. All 30 sera were negative in the Anti-nRNP/Sm ELISA (IgG), so no cross reactivity is expected.

Interference: To investigate the influence from hemoglobin, triglycerides and bilirubin, 5 different sera at different anti-nRNP/Sm concentrations (ratio 0.9 - 5.6) were spiked with potential interfering substances and were incubated with the test system acording to the package insert. Interferences from rheumatoid factor were investigated similarly by spiking with a RF positive serum. The recovery in relation to the unspiked sample without interferent was calculated. The individual recovery was within the range of 85 - 109 %. No significant interference was observed for concentrations of up to 1000 mg/dl for hemoglobin, 2000 mg/dl for triglyceride, 40 mg/dl for billrubin and 500 IU/ml for rheumatoid factor.

Assay cut-off: g.
Ratio 1.0

2. Comparison studies:

Method comparison with predicate device: a.
A comparison study was performed using 287 clinically characterized samples (52 MCTD, 69 SLE, 51 Sjögren's syndrome, 15 systemic sclerosis, 15 fibromyalgia, 35 RA, 30 borreliosis and 20 healthy), obtained from different sources from Europe and North America. The panel consisted of 49 men and 238 women. Age ranged from 14 to 85 years with an average age of 46 years. AntinRNP/Sm antibodies are expected in either MCTD or SLE. The other disease groups serve as control cohorts. The samples were tested with the EUROIMMUN Anti-nRNP/Sm ELISA (IgG) and with the Inova Quanta Lite RNP ELISA as the predicate device. Of the 8 discrepant samples, one was from a MCTD patient and the other 7 were from controls.

All samplesn = 287		Predicate ELISA
		positive	negative
EUROIMMUNAnti-nRNP/Sm ELISA (IgG)	positive	83	0
	negative	8	196
Negative agreement	$196 / 196 =$	100.0%	95% C.I.: 98.1% - 100.0%
Positive agreement	$83 / 91 =$	91.2%	95% C.I.: 83.4% - 96.1%
Overall agreement	$279 / 287 =$	97.2%	95% C.I.: 94.6% - 98.8%

b. Matrix comparison:
The usability of plasma was investigated using sample pairs each of serum and corresponding plasma (EDTA, Li-heparin, Citrate). Passing-Bablok regression was calculated for the comparison of serum to plasma. The regression equation is near the ideal correlation (intercept 0; slope 1.0) indicating equivalence of concentrations between serum and the corresponding plasma matrices. The comparison below satisfies this condition. Coefficients of determination were found to be above 0.99 and %recovery compared toserum was in the range of 91 to 120 % (serum = 100 %)

	EDTA plasma	Li-heparin plasma	Citrate plasma
n	15	15	15
Regression equation(y = plasma, x = serum)	$y = 0.46 + 0.98 x$	$y = -0.25 + 1.05 x$	$y = 0.37 + 1.04$
95% C.I. of intercept	-0.56 - 2.41	-1.54 - 1.27	-1.72 - 1.82
95% C.I. of slope	0.95 - 1.01	0.97 - 1.09	0.97 - 1.09
Coefficient of determination R²	0.9988	0.9929	0.9915
Mean %recovery	101 %	103 %	105 %
Range of %recovery	91 - 117 %	92 - 114 %	93 - 120 %

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Clinical studies:

Clinical studies were performed in cooperation with different sites. In total 1046 clinically characterized samples (65 from MCTD patients, 404 from SLE patients, 151 from myositis patients and 426 from control groups) were investigated for antibodies (IgG). With the EUROIMMUN Anti-nRNP/Sm ELISA (IgG) a prevalence of 100.0% (95% C.I.: 94.5 - 100.0%) was found in MCTD (clinical sensitivity) and a prevalence of 23.3% (95% C.I.: 19.2 - 27.7%) in SLE with a specificity of 99.3% (95% C.I.: 98.0 - 99.9%). The myositis panel was not considered for calculation of sensitivity and specificity, because anti-nRNP/Sm antibodies may occur in this disease (Tomer 1993). The results are shown in the table below. 95% C.I. are calculated by the exact method.

Clinical sensitivity: a.

No.	Panel	n	Anti-nRNP/Sm ELISA (IgG)
			positive	%	95% C.I.
1	Mixed connective tissue diseases	65	65	100.0%	94.5 - 100.0%
2	Systemic lupus erythematosus	404	94	23.3%	19.2 - 27.7%

Clinical specificity: b.

No.	Panel	n	Anti-nRNP/Sm ELISA (IgG)
			negative	%	95% C.I.
3	Polymyositis/dermatomyositis	151	143	94.7%	89.8 - 97.7%
4	Rheumatoid arthritis	164	164	100.0%	97.8 - 100.0%
5	Systemic sclerosis	81	81	100.0%	95.5 - 100.0%
6	Sjögren's syndrome	88	86	97.7%	92.0 - 99.7%
7	Other autoimmune diseases*	63	62	98.4%	91.5 - 100.0%
8	Borreliosis	30	30	100.0%	88.4 - 100.0%
	Total	577	566	98.1%	96.6 - 99.9%

*from the following groups: AIH (n = 8), Grave's disease (n = 12), Hashimoto (n = 11), celiac disease (n = 11), Diabetes Type I (n = 12)

Other clinical supportive data (when a. and b. are not applicable): C. Not applicable.
1. Clinical cut-off:

See Assay cut-off.

ട. Expected values/Reference range:
The levels of anti-nRNP/Sm antibodies (IgG) were analyzed in a panel of 200 samples from apparently healthy blood donors (120 men and 80 women with an average age of 40 y; age range: 19 - 68 y). The results are shown in the table below.

n	200
Positives	1
Negatives	199
Prevalence	0.5%
	Ratio
Lowest value	0.1
Highest value	1.3
Mean value	0.1
Std deviation	0.09

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.

Michael Locke
Signature

Dir. Regulatory Affairs Title

12 June 2013 Date

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ATTACHMENT 1

PREMARKET NOTIFICATION રાજીન્દ) SAFETY AND EFFECTIVENESS SUMMARY (as required by 21 CFR § 807.92)

A. 510(k)Number: K123261
B. Purpose for Submission: New device
C. Measurand:
- Anti-Sm autoantibodies
D. Type of Test: Qualitative enzyme immunoassay
E. Applicant:
- EUROIMMUN US INC.
F. Proprietary and Established Names: EUROIMMUN Anti-Sm ELISA (IgG)

G. Regulatory Information:

1. Regulation:
- 21 CFR 866.5110 Antinuclear antibody immunological test system
1. Classification: Class II
1. Product code: LKP
1. Panel:

Immunology Intended Use:
1. Intended use(s):

1. Indication(s) for use:
- Same as intended use.
1. Special conditions for the use statement(s):
- For prescription use only.
র্ম Special instrument requirements:

Microwell plate reader capable of measuring OD at 450nm for dual wavelength readings.

l. Device Description:

J. Substantial Equivalence Information:

Predicate device name (s): 1.
Inova Quanta Lite Sm ELISA 2. Predicate 510(k) number(s):
K922831

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Image /page/6/Picture/1 description: The image shows a black and white illustration of a ladder. The ladder has four rungs that are evenly spaced apart. The rungs are connected to two vertical rails on either side.

3. Comparison with predicate:

Similarities
Item	New Device	Predicate Device
Intended use	Detection of IgG antibodies to Sm	Same
Technology	ELISA	Same
Assay platform	96-well microtiter plates	Same
Calibration	Relative units	Same
Antigen	Purified Sm antigen	Same
Conjugate	Anti-human IgG labeled with horseradish peroxidase	Same
Substrate	TMB	Same
Reagent preparation	All reagents, calibrator and controls are ready to use,except for the wash buffer.	Same
Procedure	Sample incubation with micro-well antigen coated plate,followed by a wash step, incubation with an anti-humanIgG enzyme conjugate; wash step, incubation withsubstrate; then the addition of a stop solution andreading at 450nm.	Same
Differences
Item	New Device	Predicate Device
Assay format	Qualitative	"Semi-quantitative" (using low positive control)
Sample dilution	1:201	1:101
Calibrators and	1 calibrator	3 controls: 1 high positive, 1 low positive (used for
controls	2 controls: 1 positive, 1 negative	calculation of results), 1 negative
Wash buffer	10x concentrate	40x concentrate
Stop solution	0.5 M sulphuric acid	0.344 M sulphuric acid
Sample types	Serum or plasma (EDTA, Li-heparin, Citrate)	Serum
Reported results	Ratio	Units
Cut off level	Ratio 1.0	20 Units

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

Guidance for Industry and FDA Staff: Recommendations for Antibody (ANA) Test System Premarket (510(k)) Submissions (January 22, 2009)

L. Test Principle:

Patient samples are diluted 1:201 in sample buffer, 100 pl 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 microtier well strips are washed with wash buffer to remove unbound antibodies and 100 ul of the anti-human IgG enzyme coniugate 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 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.

Performance Characteristics (where applicable): M.

Analytical performance:
- Precision/Reproducibility: a.

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Image /page/7/Picture/0 description: The image shows a repeating pattern of black shapes against a white background. The black shapes are vertically oriented and have a rounded, almost pill-like form, with a narrower section in the middle. These shapes are evenly spaced and aligned in a row, creating a rhythmic visual sequence. The top and bottom of the image are bordered by solid black lines, which frame the repeating pattern.

Intra-assay reproducibility Anti-Sm ELISA (IgG) Ratio n = 20 Sample 2 Sample 5 Sample 6 Sample 1 Sample 3 Sample 4 Mean value (x): 5.7 3.7 2.0 1.2 0.8 0.4 5.4 - 5.8 3.5 - 3.9 1.9 – 2.0 1.2 - 1.3 0.7 - 0.9 0.4 - 0.4 Range of values: negative Expected result: positive positive positive positive negative % positive: 100% 100% 100% 100% 0% 0% 0% 100% 100% % negative: 0% 0% 0%

Inter-assay reproducibility

n = 10 x 4	Anti-Sm ELISA (IgG)Ratio
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6
Mean value (x):	5.8	3.9	1.7	1.1	0.8	0.4
Range of values:	5.3 - 7.4	3.5 - 4.6	1.5 - 2.2	1.0 - 1.2	0.6 - 1.0	0.3 - 0.5
Expected result:	positive	positive	positive	positive	negative	negative
% positive:	100%	100%	100%	100%	2.5%	0%
% negative:	0%	0%	0%	0%	97.5%	100%

Lot to lot reproducibility

		Anti-Sm ELISA (IgG)
		Ratio
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6
n	6*	6*	6*	6*	6*	6*
Mean value (x):	3.6	3.4	3.1	0.3	0.9	1.2
Range of values:	3.3 - 3.9	3.2 - 3.9	2.9 - 3.4	0.3 - 0.3	0.8 - 0.9	1.1 - 1.3
Expected result:	positive	positive	positive	negative	negative	positive
% positive:	100%	100%	100%	0%	0%	100%
% negative:	0%	0%	0%	100%	100%	0%

	Anti-Sm ELISA (IgG) Ratio
	Sample 7	Sample 8	Sample 9	Sample 10	Sample 11
n	9**	10**	10**	9**	10**
Mean value (x):	0.1	1.9	3.4	5.7	8.3
Range of values:	0.0 - 0.2	1.5 - 2.2	2.6 - 3.8	4.9 - 6.7	7.5 - 9.6
Expected result:	negative	positive	positive	positive	positive
% positive:	0%	100%	100%	100%	100%
% negative:	100%	0%	0%	0%	0%

*3 lots x 2 runs ** n lots x 1 run

Linearity/assay reportable range: b. Not applicable.

High dose Hook effect C.

Potential for a high dose Hook effect is a phenomenon that is inherent with one step "sandwich" assay designs: Very high concentrations of antigen in the patient sample bind to all available sites saturating them - on both the antibody-solid phase and the antibody-labeled conjugate and thereby prevent the "sandwich" formation. Under these conditions, the measured level of analyte may besignificantly lower than the actual level present in the sample. The two-step immunoassay design of the Anti-Sm ELISA (IgG) eliminates the adverse contribution of binding proteins, endogenous interfering substances and general matrix effects due to the extra wash step.

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Image /page/8/Picture/1 description: The image shows a black and white pattern. There are four black shapes that are evenly spaced. The shapes are connected to a black line at the bottom of the image. There is a white line at the top and bottom of the image.

d. Traceability, Stability, Expected values (controls, calibrators or methods):

A recognized standard or reference material for anti-nuclear antibodies is not available. Results of this assay are given in ratios. The reactivity of the Anti-Sm ELISA was verified using the ANA reference panel of the CDC Centers for Disease Control and Prevention, Atlanta, USA.

Limit of detection: e.
Not applicable
f. Analvtical specificity:

Cross-reactivity: The quality of the antigen coated on the plates ensures a high specificity of the ELISA. Cross reactivity was investigated using a panel of 30 sera serologically positive for antibodies against rib.P-P, nRNP/Sm, SS-A, Scl-70, Jo-1, centromeres, Rubella virus, Measles virus, Herpes simplex virus type 1 and Borrelia burgdorferi. All 30 sera were negative in the Anti-Sm ELISA (IgG), so no cross reactivity is expected.

Interference: To investigate the influence from hemoglobin, triglycerides and bilirubin, 5 different sera at different anti-Sm concentrations (ratio 0.7 ~ 3.9) were spiked with potential interfering substances and were incubated with the test system according to the package insert. Interferences from rheumatoid factor were investigated similarly by spiking with a RF positive serum. The recovery in relation to the unspiked sample without interferent was calculated. The individual recovery was within the range of 90 - 111 %. No significant interference was observed for concentrations of up to 1000 ma/dl for hemoalobin, 2000 mg/dl for trialyceride, 40 mg/dl for billrubin and 500 lU/ml for rheumatoid factor.

Assay cut-off: ਉਂ
Ratio 1.0
1. Comparison studies:
- Method comparison with predicate device: a.

A comparison study was performed using 294 clinically characterized samples (128 SLE, 51 Sjögren's syndrome, 15 systemic sclerosis, 15 fibromyalgia, 35 RA, 30 borreliosis and 20 healthy), obtained from different sources from Europe and North America. The panel consisted of 60 men and 234 women. Age ranged from 14 to 85 years with an average age of 45 years. Anti-Sm antibodies are expected in SLE. The other disease groups serve as control cohorts. The samples were tested with the EUROMMUN Anti-Sm ELISA (IqG) and with the Inova Quanta Lite Sm ELISA as the predicate device. The results are shown in the table below. All of the 7 discrepant samples negative in the EUROIMMUN test were from controls. The 5 discrepant samples positive in the EUROIMMUN test were from SLE patients.

All samplesn = 294		Predicate ELISA
		positive	negative
EUROIMMUNAnti-Sm ELISA (IgG)	positive	37	5
	negative	7	245
Negative agreement	$245 / 250 = 98.0%$	95% C.I.: 95.4%	99.3%
Positive agreement	$37 / 44 = 84.1%$	95% C.I.: 69.9%	93.4%
Overall agreement	$282 / 294 = 95.9%$	95% C.I.: 93.0%	97.9%

b. Matrix comparison:
The usability of plasma was investigated using sample pairs each of serum and corresponding plasma (EDTA, Li-heparin, Citrate). Passing-Bablok regression was calculated for the comparison of serum to plasma. The regression equation is near the ideal correlation (intercept 0) indicating equivalence of concentrations between serum and the corresponding plasma matrices. Coefficients of determination were found to be above 0.99 and %recovery compared to serum was in the range of 89 to 118 % (serum = 100 %).

	EDTA plasma	Li-heparin plasma	Citrate plasma
n	16	16	16
Regression equation(y = plasma, x = serum)	$y = 0.33 + 1.00x$	$y = 0.19 + 0.97x$	$y = 0.05 + 1.02$
95% C.I. of intercept	-0.32 - 1.24	-0.92 - 1.95	-0.69 - 3.27
95% C.I. of slope	0.96 - 1.03	0.94 - 1.01	0.99 - 1.05
Coefficient of determination R2	0.9953	0.9948	0.9931
Mean %recovery	102 %	99 %	103 %
Range of %recovery	94 - 115 %	90 - 111 %	89 - 118 %

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3. Clinical studies:

Clinical studies were performed in cooperation with different sites. In total 1036 clinically characterized samples (414 from SLE patients and 622 from control groups) were investigated for antibodies (IgG). With the EUROIMMUN Anti-Sm ELISA (IgG) a prevalence of 11.4% (95% C.I.: 8.5 - 14.8%) was found in SLE with a specificity of 99.0% (95% C.I.: 97.9 - 99.6%). The results are shown in the table below. 95% C.I. are calculated by the exact method.

a. Clinical sensitivity:

No.	Panel	n	Anti-Sm ELISA (IgG)
			positive	%	95% C.I.
1	Systemic lupus erythematosus	414	47	11.4%	8.5 - 14.8%
No.	Panel	n	negative	%	95% C.I.
2	Rheumatoid arthritis	164	164	100.0%	97.8 - 100.0%
3	Systemic sclerosis	81	81	100.0%	95.5 - 100.0%
4	Sjögren's syndrome	88	88	100.0%	95.9 - 100.0%
5	Polymyositis/dermatomyositis	151	151	100.0%	97.6 - 100.0%
6	Mixed connective tissue diseases	45	39	86.7%	73.2 - 94.9%
7	Other autoimmune diseases*	63	63	100.0%	94.3 - 100.0%
8	Borreliosis	30	30	100.0%	88.4 - 100.0%
	Total	622	616	99.0%	97.9 - 99.6%

b. Clinical specificity:

*from the following groups: AIH (n = 8), Grave's disease (n = 12), Hashimoto (n = 11), celiac disease (n = 11), Diabetes Type I (n = 12)

Other clinical supportive data (when a. and b. are not applicable): C. Not applicable.
1. Clinical cut-off:

See Assay cut-off.

ട. Expected values/Reference range:
The levels of anti-Sm antibodies (IgG) were analyzed in a panel of 200 samples from apparently healthy blood donors (120 men and 80 women with an average age of 40 y; age range: 19 - 68 y). The results are shown in the table below.

n	200
Positives	0
Negatives	200
Prevalence	0.0%
	Ratio
Lowest value	0.1
Highest value	0.3
Mean value	0.1
Std deviation	0.02

N. Proposed Labeling:

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

0. Conclusion:

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

Michael Locke
Signature

Dir. Regulatory Affairs

12 June 2013 Date

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Image /page/10/Picture/1 description: The image shows a series of four dark, circular shapes positioned between two horizontal lines. The circular shapes are evenly spaced and appear to be touching both the top and bottom lines. The overall composition is simple and graphic, with a focus on the contrast between the dark shapes and the surrounding space.

ATTACHMENT 1

PREMARKET NOTIFICATION 510(K) SAFETY AND EFFECTIVENESS SUMMARY (as required by 21 CFR § 807.92)

A. 510(k)Number: K123261
Purpose for Submission: B. New device
். Measurand:
- Anti-SS-A autoantibodies ·
D. Type of Test:
Qualitative enzyme immunoassay ய் Applicant:
- EUROIMMUN US INC.
Proprietary and Established Names: F. EUROIMMUN Anti-SS-A ELISA (IgG)

G. Regulatory Information:

Regulation: 1.
- 21 CFR 866.5100 Antinuclear antibody immunological test system
1. Classification:
Class II
1. Product code:
LLL
1. Panel:
Immunology H. Intended Use:
- Intended use(s): 1.

1. Indication(s) for use: Same as intended use.
1. Special conditions for the use statement(s): For prescription use only.
র্ব : Special instrument requirements:

Microwell plate reader capable of measuring OD at 450nm for dual wavelength readings. Device Description:

l:

Substantial Equivalence Information: J.

Predicate device name (s): 1.
- Inova Quanta Lite SS-A ELISA
1. Predicate 510(k) number(s): K922830

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Image /page/11/Picture/1 description: The image shows the number 3 followed by the text 'Comparison with predicate:'. The number 3 is on the left side of the image. The text is on the right side of the image. The text is underlined.

Similarities
Item	New Device	Predicate Device
Intended use	Detection of IgG antibodies to SS-A	Same
Technology	ELISA	Same
Assay platform	96-well microtiter plates	Same
Calibration	Relative units	Same
Antigen	Purified SS-A antigen	Same
Conjugate	Anti-human IgG labeled with horseradish peroxidase	Same
Substrate	TMB	Same
Reagent preparation	All reagents, calibrator and controls are ready to use,except for the wash buffer.	Same
Procedure	Sample incubation with micro-well antigen coated plate,followed by a wash step, incubation with an anti-humanIgG enzyme conjugate; wash step, incubation withsubstrate; then the addition of a stop solution andreading at 450nm.	Same
Differences
Item	New Device	Predicate Device
Assay format	Qualitative	"Semi-quantitative" (using low positive control)
Sample dilution	1:201	1:101
Calibrators and	1 calibrator	3 controls: 1 high positive, 1 low positive (used for
controls	2 controls: 1 positive, 1 negative	calculation of results), 1 negative
Wash buffer	10x concentrate	40x concentrate
Stop solution	0.5 M sulphuric acid	0.344 M sulphuric acid
Sample types	Serum or plasma (EDTA, Li-heparin, Citrate)	Serum
Reported results	Ratio	Units
Cut off level	Ratio 1.0	20 Units

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

Guidance for Industry and FDA Staff: Recommendations for Antibody (ANA) Test System Premarket (510(k)) Submissions (January 22, 2009)

ட. Test Principle:

Patient samples are diluted 1:201 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 µl 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 ul of wash buffer to remove any unbound enzyme conjugate and 100 µl of the chromogen substrate is added. The strips are incubated for 15 minutes at room temperature and 100 µl 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.

Performance Characteristics (where applicable): M.

Analytical performance:
Precision/Reproducibility: ·
- The reproducibility of the test was investigated using sera with different concentrations. Intra-assay reproducibility is based on 20 determinations and inter-assay reproducibility on 40 determinations performed in 10 different runs on 5 days with 2 runs per day, each run performed according to the package insert. The following results were obtained:

Image /page/11/Picture/13 description: The image shows a pattern of alternating black and white shapes. The black shapes are rectangular bars, and the white shapes are oval-like shapes. The black bars are arranged horizontally, and the white shapes are arranged vertically between the bars. There are four white shapes in the image.

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Image /page/12/Picture/0 description: The image shows a pattern of black shapes against a white background. The shapes are arranged in a row, with each shape consisting of two horizontal lines connected by a rounded, oval-like form in the middle. The pattern is symmetrical and evenly spaced, creating a repeating design.

Intra-assay reproducibility

n = 20	Anti-SS-A ELISA (IgG)Ratio
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6
Mean value (x):	6.1	4.2	2.0	1.3	0.8	0.3
Range of values:	5.9 - 6.4	4.0 - 4.5	1.9 - 2.1	1.2 - 1.4	0.7 - 0.8	0.3 - 0.3
Expected result:	positive	positive	positive	positive	negative	negative
% positive:	100%	100%	100%	100%	0%	0%
% negative:	0%	0%	0%	0%	100%	100%

Inter-assay reproducibility

n = 10 x 4	Anti-SS-A ELISA (IgG)Ratio
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6
Mean value (x):	6.0	4.0	1.8	1.2	0.8	0.3
Range of values:	5.0 - 6.4	3.7 - 4.5	1.6 - 1.9	1.0 - 1.4	0.7 - 0.9	0.3 - 0.4
Expected result:	positive	positive	positive	positive	negative	negative
% positive:	100%	100%	100%	100%	0%	0%
% negative:	0%	0%	0%	0%	100%	100%

Lot to lot reproducibility

	Anti-SS-A ELISA (IgG)Ratio
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6
n	6*	6*	6*	6*	6*	6*
Mean value (x):	4.6	5.6	5.6	0.3	0.9	1.1
Range of values:	4.4 - 4.8	5.1 - 6.2	5.1 - 6.0	0.3 - 0.4	0.9 - 0.9	1.1 - 1.2
Expected result:	positive	positive	positive	negative	negative	positive
% positive:	100%	100%	100%	0%	0%	100%
% negative:	0%	0%	0%	100%	100%	0%

	Anti-SS-A ELISA (IgG)Ratio
	Sample 7	Sample 8	Sample 9	Sample 10	Sample 11
n	10**	11**	10**	9**	11**
Mean value (x):	0.1	1.6	2.7	4.6	7.8
Range of values:	0.1 - 0.2	1.3 - 1.8	2.3 - 3.1	4.3 - 5.2	7.4 - 8.3
Expected result:	negative	positive	positive	positive	positive
% positive:	0%	100%	100%	100%	100%
% negative:	100%	0%	0%	0%	0%

*3 lots x 2 runs ** n lots x 1 run

b. Linearity/assay reportable range: Not applicable.

Hiah dose Hook effect C.

Potential for a high dose Hook effect is a phenomenon that is inherent with one step "sandwich" assay designs: Very high concentrations of antigen in the patient sample bind to all available sites saturating them - on both the antibody-solid phase and the antibody-labeled conjugate and thereby prevent the "sandwich" formation. Under these conditions, the measured level of analyte may be significantly lower than the actual level present in the sample. The two-step immunoassay design of the Anti-SS-A ELISA (IgG) eliminates the adverse contribution of binding proteins, endogenous interfering substances and general matrix effects due to the extra wash step.

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Image /page/13/Picture/1 description: The image shows a pattern of alternating black and white shapes. There are four black, oval-like shapes evenly spaced between two horizontal black lines. The white space between the black shapes and lines creates a contrasting pattern.

Traceability, Stability, Expected values (controls, calibrators or methods): d.

A recognized standard or reference material for anti-nuclear antibodies is not available. Results of this assay are given in ratios. The reactivity of the Anti-SS-A ELISA was verified using the ANA reference panel of the CDC Centers for Disease Control and Prevention, Atlanta, USA.

e. Limit of detection:
Not applicable.
f. Analytical specificity:

Cross-reactivity: The quality of the antigen coated on the plates ensures a high specificity of the ELISA. Cross reactivity was investigated using a panel of 30 sera serologically positive for antibodies against rib.P-P, nRNP/Sm, Sm, Scl-70, Jo-1, centromeres, Rubella virus, Measles virus, Herpes simplex virus type 1 and Borrelia burgdorferi. All 30 sera were negative in the Anti-SS-A ELISA (IgG), so no cross reactivity is expected.

Interference: To investigate the influence from hemoglobin, triglycerides and bilirubin, 5 different sera at different anti-SS-A concentrations (15 - 124 RU/ml) were spiked with potential interfering substances and were incubated with the test system according to the package insert. Interferences from rheumatoid factor were investigated similarly by spiking with a RF positive serum. The recovery in relation to the unspiked sample without interferent was calculated. The individual recovery was within the range of 93 - 107 %. No significant interference was observed for concentrations of up to 1000 mg/dl for hemoglobin, 2000 mg/dl for triglyceride, 40 mg/dl for billirubin and 500 IU/ml for rheumatoid factor.

ದ್ರ. Assay cut-off:
. .

Ratio 1.0

2. Comparison studies:

Method comparison with predicate device: a.
A comparison study was performed using 305 clinically characterized samples (63 SLE, 77 Sjögren's syndrome, 23 systemic sclerosis, 1 SSc/SS, 15 fibromyalgia, 26 myositis, 35 RA, 30 borreliosis, 20 healthy), obtained from different sources from Europe and North America. The panel consisted of 54 men and 251 women. Age ranged from 14 to 85 years with an average age of 48 years. Anti-SS-A antibodies are expected in either Siögren's syndrome or SLE. The other disease groups serve as control cohorts. The samples were tested with the EUROIMMUN Anti-SS-A ELISA (IgG) and with the Inova Quanta Lite SS-A ELISA as the predicate device. Of the 9 discrepant samples negative in the EUROIMMUN test. 2 were from patients with Siögren's syndrome and one from a SLE patient, the other 6 were from controls. All 3 discrepant samples positive in the EUROIMMUN test were from SLE patients.

All samplesn = 305		Predicate ELISA
		positive	negative
EUROIMMUNAnti-SS-A ELISA (IgG)	positive	116	3
	negative	9	177
Negative agreement 177 / 180 = 98.3% 95% C.I.: 95.2% - 99.7%
Positive agreement 116 / 125 = 92.8% 95% C.I.: 86.8% - 96.7%
Overall agreement 293 / 305 = 96.1% 95% C.I.: 93.2% - 98.0%

b. Matrix comparison:
The usability of plasma was investigated using sample pairs each of serum and corresponding plasma (EDTA, Li-heparin, Citrate). Passing-Bablok regression was calculated for the comparison of serum to plasma. The regression equation is near the ideal correlation (intercept 0; slope 1.0) indicating equivalence of concentrations between serum and the corresponding plasma matrices. Coefficients of determination were found to be above 0.99 and %recovery compared to serum was in the range of 85 to 104 % (serum = 100 %).

	EDTA plasma	Li-heparin plasma	Citrate plasma
n	16	16	16
Regression equation(y = plasma, x = serum)	$y = -1.59 + 0.99 x$	$y = -1.47 + 0.99 x$	$y = -0.54 + 0.98$
95% C.I. of intercept	-4.49 - 3.01	-4.62 - 1.36	-4.65 - 3.32
95% C.I. of slope	0.95 - 1.02	0.95 - 1.02	0.94 - 1.04
Coefficient of determination R²	0.9972	0.9929	0.9956
Mean %recovery	97 %	95 %	98 %
Range of %recovery	90 - 102 %	85 - 102 %	93 - 104 %

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Clinical studies:

Clinical studies were performed in cooperation with different sites. In total 1026 clinically characterized samples (88 from SS patients, 404 from SLE patients and 534 from control groups) were investigated for anti-SS-A antibodies (IgG). With the EUROIMMUN Anti-SS-A ELISA (IgG) a prevalence of 73.9% (95% C.I.: 63.4 – 82.7%) was found in Sjögren's syndrome (clinical sensitivity) and a prevalence of 40.6% (95% C.I.: 35.8 - 45.6%) was found in SLE with a specificity of 94.8% (95% C.I.: 92.5 - 96.5%). Systemic sclerosis and myositis panels were considered for calculation of specificity, however, it has been reported that anti-SS-A antibodies may occur in these diseases [Antonioli 2002, Ghirardello 2005]. The results are shown in the table below. 95% C.I. are calculated by the exact method.

Clinical sensitivity: a.

No.	Panel	n	positive	%	95% C.I.
1	Sjögren's syndrome	88	65	73.9%	63.4 - 82.7%
2	Systemic lupus erythematosus	404	164	40.6%	35.8 - 45.6%

Clinical specificity: b.

No.	Panel	n	negative	%	95% C.I.
3	Systemic sclerosis	81	75	92.6%	84.6 - 97.2%
4	Polymyositis/dermatomyositis	151	138	91.4%	85.7 - 95.3%
5	Rheumatoid arthritis	164	159	97.0%	93.0 - 99.0%
6	Mixed connective tissue diseases	45	41	91.1%	78.8 - 97.5%
7	Other autoimmune diseases*	63	63	100.0%	94.3 - 100.0%
8	Borreliosis	30	30	100.0%	88.4 - 100.0%
	Total	534	506	94.8%	92.5 - 96.5%

ffrom the following groups: AIH (n = 8), Grave's disease (n = 12), Hashimoto (n = 11), celiac disease (n = 11), Diabetes Type I (n = 12)

Other clinical supportive data (when a. and b. are not applicable). C. Not applicable.
1. Clinical cut-off:

See Assay cut-off.

1. Expected values/Reference range:
  The levels of anti-SS-A antibodies (IgG) were analyzed in a panel of 200 samples from apparently healthy blood donors (120 men and 80 women with an average age of 40 y; age range: 19 - 68 y). The results are shown in the table below.

n	200
Positives	2
Negatives	198
Prevalence	1.0%
	Ratio
Lowest value	0.0
Highest value	4.4
Mean value	0.1
Std deviation	0.33

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.

Michael Locke

Signature

Dir. Regulatory Affairs Title

12 June 2013 Date

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ATTACHMENT 1

PREMARKET NOTIFICATION 210(K) SAFETY AND EFFECTIVENESS SUMMARY (as required by 21 CFR § 807.92)

A. 510(k)Number: K123261
Purpose for Submission: в. New device
C. Measurand:
- Anti-SS-B autoantibodies
D. Type of Test:
- Qualitative enzyme immunoassay
E. Applicant:
- EUROIMMUN US INC.
F. Proprietary and Established Names: EUROIMMUN Anti-SS-B ELISA (IqG)

Regulatory Information: G.

1. Regulation:
- 21 CFR 866.5110 Antinuclear antibody immunological test system
1. Classification: Class II
1. Product code: LLL
র্ব Panel:
Immunology
H. Intended Use:
- Intended use(s): 1.

1. Indication(s) for use: Same as intended use.
ന Special conditions for the use statement(s): For prescription use only .

Special instrument requirements: 4.

Microwell plate reader capable of measuring OD at 450nm for dual wavelength readings. Device Description:

l.

Substantial Equivalence Information: J.

Predicate device name (s): 1.
Inova Quanta Lite SS-B ELISA
1. Predicate 510(k) number(s): K922832

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റ്റ് Comparison with predicate:

Similarities
Item	New Device	Predicate Device
Intended use	Detection of IgG antibodies to SS-B	Same
Technology	ELISA	Same
Assay platform	96-well microtiter plates	Same
Calibration	Relative units	Same
Antigen	Purified SS-B antigen	Same
Conjugate	Anti-human IgG labeled with horseradish peroxidase	Same
Substrate	TMB	Same
Reagent preparation	All reagents, calibrator and controls are ready to use,except for the wash buffer.	Same
Procedure	Sample incubation with micro-well antigen coated plate,followed by a wash step, incubation with an anti-humanIgG enzyme conjugate; wash step, incubation withsubstrate; then the addition of a stop solution andreading at 450nm.	Same
Differences
Item	New Device	Predicate Device
Assay format	Qualitative	"Semi-quantitative" (using low positive control)
Sample dilution	1:201	1:101
Calibrators and	1 calibrator	3 controls: 1 high positive, 1 low positive (used for
controls	2 controls: 1 positive, 1 negative	calculation of results), 1 negative
Wash buffer	10x concentrate	40x concentrate
Stop solution	0.5 M sulphuric acid	0.344 M sulphuric acid
Sample types	Serum or plasma (EDTA, Li-heparin, Citrate)	Serum
Reported results	Ratio	Units
Cut off level	Ratio 1.0	20 Units

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

Guidance for Industry and FDA Staff: Recommendations for Antibody (ANA) Test System Premarket (510(k)) Submissions (January 22, 2009)

L. Test Principle:

Patient samples are diluted 1:201 in sample buffer, 100 pl 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 ul of wash buffer to remove any unbound enzyme conjugate and 100 µl of 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.

Performance Characteristics (where applicable): M.

Analytical performance: 1 -
- Precision/Reproducibility: a.

The reproducibility of the test was investigated using sera with different concentrations. Intra-assay reproducibility is based on 20 determinations and inter-assay reproducibility on 40 determinations performed in 10 different runs on 5 days with 2 runs per day, each run performed according to the package insert. The following results were obtained:

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Image /page/17/Picture/0 description: The image shows a close-up of a black and white pattern. The pattern consists of a series of vertical black bars that are connected by horizontal black lines at the top and bottom. The spaces between the vertical bars are white and have a rounded, oval shape. There are four of these oval shapes visible in the image.

Intra-assay reproducibility
			Anti-SS-B ELISA (IgG)
n = 20			Ratio
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6
Mean value (x):	4.1	2.9	1.3	0.8	0.6	0.4
Range of values:	4.0 - 4.3	2.8 - 3.1	1.1 - 1.4	0.7 - 0.9	0.5 - 0.7	0.4 - 0.5
Expected result:	positive	positive	positive	negative	negative	negative
% positive:	100%	100%	100%	0%	0%	0%
% negative:	0%	0%	0%	100%	100%	100%

Inter-assay reproducibility

n = 10 x 4	Anti-SS-B ELISA (IgG)Ratio
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6
Mean value (x):	4.7	3.2	1.5	1.1	0.7	0.5
Range of values:	4.2 - 5.3	3.0 - 3.7	1.3 - 1.7	1.0 - 1.1	0.5 - 0.7	0.4 - 0.5
Expected result:	positive	positive	positive	positive	negative	negative
% positive:	100%	100%	100%	100%	0%	0%
% negative:	0%	0%	0%	0%	100%	100%

Lot to lot reproducibility

	Anti-SS-B ELISA (IgG) Ratio
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6
n	6*	6*	6*	6*	6*	6*
Mean value (x):	3.9	3.9	3.8	0.3	0.9	1.2
Range of values:	3.6 - 4.2	3.4 - 4.4	3.3 - 4.1	0.3 - 0.3	0.9 - 0.9	1.1 - 1.3
Expected result:	positive	positive	positive	negative	negative	positive
% positive:	100%	100%	100%	0%	0%	100%
% negative:	0%	0%	0%	100%	100%	0%

	Anti-SS-B ELISA (IgG) Ratio
	Sample 7	Sample 8	Sample 9
n	11**	11**	11**
Mean value (x):	2.6	4.2	7.7
Range of values:	2.3 - 3.1	3.2 - 4.8	7.0 - 8.8
Expected result:	positive	positive	positive
% positive:	100%	100%	100%
% negative:	0%	0%	0%

*3 lots x 2 runs ** n lots x 1 run

b. Linearity/assay reportable range: Not applicable.

High dose Hook effect C.

Potential for a high dose Hook effect is a phenomenon that is inherent with one step "sandwich" assay designs: Very high concentrations of antigen in the patient sample bind to all available sites saturating them - on both the antibody-solid phase and the antibody-labeled conjugate and thereby prevent the "sandwich" formation. Under these conditions, the measured level of analyte may be significantly lower than the actual level present in the sample. The two-step immunoassay design of the Anti-SS-B ELISA (IgG) eliminates the adverse contribution of binding proteins, endogenous interfering substances and general matrix effects due to the extra wash step.

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Image /page/18/Picture/1 description: The image shows a pattern of four black circles evenly spaced between two horizontal black lines. The circles are solid black and appear to be the same size and shape. The lines are also solid black and appear to be parallel to each other. The circles are positioned in the center between the two lines. The pattern is simple and symmetrical.

d. Traceability, Stability, Expected values (controls, calibrators or methods):

A recognized standard or reference material for anti-nuclear antibodies is not available. Results of this assay are given in ratios. The reactivity of the Anti-SS-B ELISA was verified using the ANA reference panel of the CDC Centers for Disease Control and Prevention, Atlanta, USA.

Limit of detection: e.
Not applicable.
f. Analytical specificity:

Cross-reactivity: The quality of the antigen coated on the plates ensures a high specificity of the ELISA. Cross reactivity was investigated using a panel of 30 sera serologically positive for antibodies against rib.P-P, nRNP/Sm, Sm. SS-A, Scl-70, Jo-1, centromeres, Rubella virus, Measles virus, Herpes simplex virus type 1 and Borrelia burgdorferi. All 30 sera were negative in the Anti-SS-B ELISA (IgG), so no cross reactivity is expected.

Interference: To investigate the influence from hemoglobin, triglycerides and bilirubin, 5 different sera at different anti-SS-B concentrations (18 - 123 RU/ml) were spiked with potential interfering substances and were incubated with the test system according to the package insert. Interferences from rheumatoid factor were investigated similarly by spiking with a RF positive serum. The recovery in relation to the unspiked sample without interferent was calculated. The individual recovery was within the range of 92 - 116 %. No significant interference was observed for concentrations of up to 1000 mg/dl for hemoglobin, 2000 mg/dl for triglyceride, 40 mg/dl for billrubin and 500 IU/ml for rheumatoid factor.

Assay cut-off: টা
Ratio 1.0
1. Comparison studies:
- Method comparison with predicate device: a.

A comparison study was performed using 275 clinically characterized samples (57 SLE, 64 Sjögren's syndrome, 23 systemic sclerosis, 1 SSc/SS, 4 SLE/SS, 15 fibromyalgia, 26 myositis, 35 RA, 30 borreliosis and 20 healthy), obtained from different sources from Europe and North America. The panel consisted of 55 men and 220 women. Age ranged from 12 to 83 years with an average age of 48 vears. Anti-SS-B antibodies are expected in either Sjögren's syndrome or SLE. The other disease groups serve as control cohorts. The samples were tested with the EUROIMMUN Anti-SS-B ELISA (IgG) and with the Inova Quanta Lite SS-B ELISA as the predicate device. Of the 3 discrepant samples negative in the EUROIMMUN test, one was from a Siögren's syndrome patient and the other 2 were from controls. The discrepant sample positive in the EUROIMMUN test was from a Sjögren's syndrome patient.

All samplesn = 275		Predicate ELISA
		positive	negative
EUROIMMUNAnti-SS-B ELISA (IgG)	positive	82	1
	negative	3	189
Negative agreement		$189 / 190 = 99.5%$	95% C.I.: 97.1% - 100.0%
Positive agreement		$82 / 85 = 96.5%$	95% C.I.: 90.0% - 99.3%
Overall agreement		$271 / 275 = 98.5%$	95% C.I.: 96.3% - 99.6%

b. Matrix comparison:
The usability of plasma was investigated using sample pairs each of serum and corresponding plasma (EDTA, Li-heparin, Citrate). Passing-Bablok regression was calculated for the comparison of serum to plasma. The regression equation is near the ideal correlation (intercept 0; slope 1.0) indicating equivalence of concentrations between serum and the corresponding plasma matrices. Coefficients of determination were found to be above 0.99 and %recovery compared toserum was in the range of 78 to 116 % (serum = 100 %).

	EDTA plasma	Li-heparin plasma	Citrate plasma
n	16	16	16
Regression equation(y = plasma, x = serum)	$y = -1.56 + 1.05 x$	$y = -0.22 + 1.06 x$	$y = 0.55 + 1.01$
95% C.I. of intercept	-5.46 - 0.19	-3.29 - 2.28	-1.56 - 5.62
95% C.I. of slope	1.04 - 1.09	1.01 - 1.09	0.97 - 1.05
Coefficient of determination R2	0.9972	0.9968	0.9951
Mean %recovery	100 %	105 %	101 %
Range of %recovery	78 - 116 %	98 - 116 %	81 - 110 %

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Image /page/19/Picture/1 description: The image shows a repeating pattern of black shapes against a white background. The pattern consists of a horizontal line with rounded shapes above and below it. The rounded shapes are evenly spaced and appear to be identical.

Clinical studies: ന്
Clinical studies were performed in cooperation with different sites. In total 1026 clinically characterized samples (88 from SS patients, 404 from SLE patients and 534 from control groups) were investigated for anti-SS-B antibodies (IgG). With the EUROIMMUN Anti-SS-B ELISA (IgG) a prevalence of 39.8% (95% C.I.: 29.5 - 50.8%) was found in Siggren's syndrome and a prevalence of 13.9% (95% C.I.: 10.6 - 17.6%) was found in SLE with a specificity of 98.1% (95% C.I.: 96.6 – 99.1%). The results are shown in the table below. 95% C.I. are calculated by the exact method.
Clinical sensitivity: a.

No.	Panel	n	Anti-SS-B ELISA (IgG)
			positive	%	95% C.I.
1	Sjögren's syndrome	88	35	39.8%	29.5 – 50.8%
2	Systemic lupus erythematosus	404	56	13.9%	10.6 – 17.6%

Clinical specificity: b.

No.	Panel	n	Anti-SS-B ELISA (IgG)
			negative	%	95% C.I.
3	Rheumatoid arthritis	164	163	99.4%	96.6 - 100.0%
4	Systemic sclerosis	81	77	95.1%	87.8 - 98.6%
5	Polymyositis/dermatomyositis	151	149	98.7%	95.3 - 99.8%
6	Mixed connective tissue diseases	45	42	93.3%	81.7 - 98.6%
7	Other autoimmune diseases*	63	63	100.0%	94.3 - 100.0%
8	Borreliosis	30	30	100.0%	88.4 - 100.0%
	Total	534	524	98.1%	96.6 - 99.1%

from the following groups: AIH (n = 8), Grave's disease (n = 12), Hashimoto (n = 11), celiac disease (n = 11), Diabetes Type I (n = 12)

Other clinical supportive data (when a. and b. are not applicable): C. Not applicable.
1. Clinical cut-off:

See Assay cut-off.

Expected values/Reference range: 5.
The levels of anti-SS-B antibodies (IgG) were analyzed in a panel of 200 samples from apparently healthy blood donors (120 men and 80 women with an average age of 40 y; age range: 19 - 68 y). The results are shown in the table below.

n	200
Positives	0
Negatives	200
Prevalence	0.0%
	Ratio
Lowest value	0.0
Highest value	0.2
Mean value	0.0
Std deviation	0.02

N. Proposed Labeling:

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

். Conclusion:

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

Michael Locke
Signature

Dir. Regulatory Affairs Title

12 June 2013 Date

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Image /page/20/Figure/1 description: The image shows a pattern of alternating black and white shapes. The black shapes are oval or circular, and they are arranged in a row. The white shapes are rectangular, and they are arranged in a row above and below the black shapes. The pattern is symmetrical and repetitive.

ATTACHMENT I

PREMARKET NOTIFICATION 510(K) SAFETY AND EFFECTIVENESS SUMMARY (as required by 21 CFR § 807.92)

A. 510(k)Number: · K123261
Purpose for Submission: B. New device
C. Measurand:
- Anti-Scl-70 autoantibodies
D. Type of Test: Qualitative enzyme immunoassay
E. Applicant:
- EUROIMMUN US INC.
F. Proprietary and Established Names: EUROIMMUN Anti-Scl-70 ELISA (IgG)
G. Requlatory Information:
- Regulation: 1.
  - 21 CFR 866.5110 Antinuclear antibody immunological test system
- 1. Classification: Class II
- 1. Product code: LLL
- র . Panel: Immunology
H. Intended Use:
- 1. Intended use(s):

1. Indication(s) for use: 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. Device Description:

l.
The EUROIMMUN Anti-Scl-70 ELISA (IgG) consists of a microwell ELISA plate coated with Scl-70 antigen, calibrator, positive and negative control, peroxidase-labelled anti-human IgG conjugate, sample buffer, wash buffer concentrate, TMB chromogen/substrate solution and stop solution.

J. Substantial Equivalence Information:

Predicate device name (s): 1.
- Inova Quanta Lite Scl-70 ELISA
1. Predicate 510(k) number(s): K924898

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ന് Comparison with predicate:

SimilaritiesItem	New Device	Predicate Device
Intended use	Detection of IgG antibodies to Scl-70	Same
Technology	ELISA	Same
Assay platform	96-well microtiter plates	Same
Calibration	Relative units	Same
Antigen	Purified Scl-70 antigen	Same
Conjugate	Anti-human IgG labeled with horseradish peroxidase	Same
Substrate	TMB	Same
Reagent preparation	All reagents, calibrator and controls are ready to use,except for the wash buffer.	Same
Procedure	Sample incubation with micro-well antigen coated plate,followed by a wash step, incubation with an anti-humanIgG enzyme conjugate; wash step, incubation withsubstrate; then the addition of a stop solution andreading at 450nm.	Same
Differences
Item	New Device	Predicate Device
Assay format	Qualitative	"Semi-quantitative" (using low positive control)
Sample dilution	1:201	1:101
Calibrators and controls	1 calibrator2 controls: 1 positive, 1 negative	3 controls: 1 high positive, 1 low positive (used forcalculation of results), 1 negative
Wash buffer	10x concentrate	40x concentrate
Stop solution	0.5 M sulphuric acid	0.344 M sulphuric acid
Sample types	Serum or plasma (EDTA, Li-heparin, Citrate)	Serum
Reported results	Ratio	Units

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

Guidance for Industry and FDA Staff: Recommendations for Antibody (ANA) Test System Premarket (510(k)) Submissions (January 22, 2009)

20 Units

L. Test Principle:

Cut off level

Patient samples are diluted 1:201 in sample buffer. 100 ul 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 microfiter wells are again washed 3 times with 300 µl of wash buffer to remove any unbound enzyme conjugate and 100 µl of 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. Performance Characteristics (where applicable):

Analytical performance: 1.

a. Precision/Reproducibility:
Ratio 1.0

The reproducibility of the test was investigated using sera with different concentrations. Intra-assay reproducibility is based on 20 determinations and inter-assay reproducibility on 40 determinations performed in 10 different runs on 5 days with 2 runs per day, each run performed according to the package insert. The following results were obtained:

{22}------------------------------------------------

Intra-assay reproducibility
	Anti-Scl-70 ELISA (IgG)
n = 20
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6
Mean value (x):	6.4	4.2	2.1	1.4	0.8	0.4
Range of values:	5.8 - 6.7	3.8 - 4.5	2.0 - 2.3	1.3 - 1.5	0.7 - 0.9	0.3 - 0.4
Expected result:	positive	positive	positive	positive	negative	negative
% positive:	100%	100%	100%	100%	0%	0%
% negative:	0%	0%	0%	0%	100%	100%

Inter-assay reproducibility

n = 10 x 4	Anti-Scl-70 ELISA (IgG)Ratio
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6
Mean value (x):	6.2	4.2	1.9	1.2	0.8	0.3
Range of values:	5.7 - 7.0	3.7 - 4.7	1.7 - 2.2	1.1 - 1.4	0.7 - 0.9	0.2 - 0.4
Expected result:	positive	positive	positive	positive	negative	negative
% positive:	100%	100%	100%	100%	0%	0%
% negative:	0%	0%	0%	0%	100%	100%

Lot to lot reproducibility

	Anti-Scl-70 ELISA (IgG)
	Ratio
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6
n	6*	6*	6*	6*	6*	6*
Mean value (x):	4.5	4.7	3.6	0.3	0.9	1.1
Range of values:	4.0 - 5.0	4.1 - 5.4	3.3 - 3.8	0.3-0.3	0.8 - 0.9	1.0 - 1.2
Expected result:	positive	positive	positive	negative	negative	positive
% positive:	100%	100%	100%	0%	0%	100%
% negative:	0%	0%	0%	100%	100%	0%

		Anti-Scl-70 ELISA (IgG)Ratio
	Sample 7	Sample 8	Sample 9
n	11**	10**	11**
Mean value (x):	3.0	4.8	6.2
Range of values:	2.5 - 3.7	4.3 - 5.7	5.3 - 6.8
Expected result:	negative	positive	positive
% positive:	0%	100%	100%
% negative:	100%	0%	0%

*3 lots x 2 runs ** n lots x 1 run

b. Linearity/assay reportable range: Not applicable.

High dose Hook effect C.

Potential for a high dose Hook effect is a phenomenon that is inherent with one step "sandwich" assay designs: Very high concentrations of antigen in the patient sample bind to all available sites saturating them - on both the antibody-solid phase and the antibody-labeled conjugate and thereby prevent the "sandwich" formation. Under these conditions, the measured level of analyte may be significantly lower than the actual level present in the sample. The two-step immunoassay design of the Anti-Scl-70 ELISA (IgG) eliminates the adverse contribution of binding proteins, endogenous interfering substances and general matrix effects due to the extra wash step.

{23}------------------------------------------------

Image /page/23/Picture/1 description: The image shows a black and white pattern of alternating dark and light vertical bars. The dark bars are wider in the middle and narrower at the top and bottom, creating a rounded shape. The light bars are uniform in width and separate the dark bars. The pattern is repeated several times across the image.

d. Traceability. Stability, Expected values (controls, calibrators or methods):

A recognized standard or reference material for anti-nuclear antibodies is not available. Results of this assay are given in ratios. The reactivity of the Anti-Scl-70 ELISA was verified using the ANA reference panel of the CDC Centers for Disease Control and Prevention, Atlanta, USA.

Limit of detection: e.
Not applicable.
f. Analytical specificity:

Cross-reactivity: The quality of the antigen coated on the plates ensures a high specificity of the ELISA. Cross reactivity was investigated using a panel of 30 sera serologically positive for antibodies aqainst rib.P-P, nRNP/Sm, Sm, SS-A, SS-B, Jo-1, centromeres, Rubella virus, Measles virus, Herpes simplex virus type 1 and Borrelia burgdorferi. All 30 sera were negative in the Anti-Scl-70 ELISA (IgG), so no cross reactivity is expected.

Interference: To investigate the influence from hemoglobin, triglycerides and bilirubin, 5 different sera at different anti-Scl-70 concentrations (17 - 100 RU/ml) were spiked with potential interfering substances and were incubated with the test system according to the package insert. Interferences from rheumatoid factor were investigated similarly by spiking with a RF positive serum. The recovery in relation to the unspiked sample without interferent was calculated. The individual recovery was within the range of 86 - 109 %. No significant interference was observed for concentrations of up to 1000 mg/dl for hemoglobin, 2000 mg/dl for triglyceride, 40 mg/dl for bilirubin and 500 lU/ml for rheumatoid factor.

Assay cut-off:
Ratio 1.0

ರು

2. Comparison studies:

Method comparison with predicate device: a.
A comparison study was performed using 309 clinically characterized samples (158 systemic sclerosis, 51 Sjögren's syndrome, 15 fibromyalgia, 35 RA, 30 borreliosis and 20 healthy), obtained from different sources from Europe and North America. The panel consisted of 62 men and 247 women. Age ranged from 14 to 88 years with an average age of 53 years. Anti-Scl-70 antibodies are expected in systemic sclerosis. The other disease groups serve as control cohorts. The samples were tested with the EUROIMMUN Anti-Scl-70 ELISA (IgG) and with the Inova Quanta Lite Scl-70 ELISA as the predicate device.

All samplesn = 309		Predicate ELISA
		positive	negative
EUROIMMUNAnti-Scl-70 ELISA (IgG)	positive	125	0
EUROIMMUNAnti-Scl-70 ELISA (IgG)	negative	0	184
Negative agreement	$184 / 184 = 100.0%$	95% C.I.: 98.0%	100.0%
Positive agreement	$125 / 125 = 100.0%$	95% C.I.: 97.1%	100.0%
Overall agreement	$309 / 309 = 100.0%$	95% C.I.: 98.8%	100.0%

b. Matrix comparison:
The usability of plasma was investigated using sample pairs each of serum and corresponding plasma (EDTA, Li-heparin, Citrate). Passing-Bablok regression was calculated for the comparison of serum to plasma. The regression equation is near the ideal correlation (intercept 0; slope 1.0) indicating equivalence of concentrations between serum and the corresponding plasma matrices. Coefficients of determination were found to be above 0.98 and %recovery compared toserum was in the range of 86 to 113 % (serum = 100 %).

	EDTA plasma	Li-heparin plasma	Citrate plasma
n	16	16	16
Regression equation(y = plasma, x = serum)	$y = -0.58 + 1.00 x$	$y = 0.17 + 1.02 x$	$y = -0.87 + 0.97$
95% C.I. of intercept	-3.25 - 2.24	-1.99 - 1.64	-2.02 - 1.35
95% C.I. of slope	0.94 - 1.06	0.98 - 1.06	0.94 - 1.03
Coefficient of determination R2	0.9895	0.9930	0.9953
Mean %recovery	98 %	101 %	96 %
Range of %recovery	86 - 107 %	92 - 113 %	87 - 103 %

{24}------------------------------------------------

3. Clinical studies:

Clinical studies were performed in cooperation with different sites. In total 909 clinically characterized samples (280 from systemic sclerosis patients and 629 from control groups) were investigated for anti-Scl-70 antibodies (IgG). With the EUROMMUN Anti-Scl-70 ELISA (IgG) a prevalence of 23.2% (95% C.I.: 18.4 - 28.6%) was found in systemic sclerosis, a prevalence of 59.4% (95% C.I.: 48.9 - 69.3%) was found in diffuse systemic sclerosis and a prevalence of 5.3% (95% C.I.: 2.0 - 11.2%) was found in limited systemic sclerosis with a specificity of 99.8% (95% C.I.: 99.1 - 100.0%).The results are shown in the table below. 95% C.I. are calculated by the exact method.

a. Clinical sensitivity:

No.	Panel	n	positive	%	95% C.I.
1	Systemic sclerosis	280	65	23.2%	18.4 – 28.6%
1a	Diffuse systemic sclerosis (from panel 1)	96	57	59.4%	48.9 – 69.3%
1b	Limited systemic sclerosis (from panel 1)	113	6	5.3%	2.0 – 11.2%

b. Clinical specificity:

No.	Panel	n	negative	%	95% C.I.
2	Systemic lupus erythematosus	213	213	100.0%	98.3 - 100.0%
3	Polymyositis/dermatomyositis	26	26	100.0%	86.8 - 100.0%
4	Rheumatoid arthritis	164	163	99.4%	96.6 - 100.0%
5	Sjögren's syndrome	88	88	100.0%	95.9 - 100.0%
6	Mixed connective tissue diseases	45	45	100.0%	92.1 - 100.0%
7	Other autoimmune diseases*	63	63	100.0%	94.3 - 100.0%
8	Borreliosis	30	30	100.0%	88.4 - 100.0%
	Total	629	628	99.8%	99.1 - 100.0%

ffrom the following groups: AIH (n = 8), Grave's disease (n = 12), Hashimoto (n = 11), celiac disease (n = 11), Diabetes Type I (n = 12)

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

র্ব : Clinical cut-off:

See Assay cut-off.

5. Expected values/Reference range:

The levels of anti-Scl-70 antibodies (IgG) were analyzed in a panel of 200 samples from apparently healthy blood donors (120 men and 80 women with an average age of 40 y; age range: 19 - 68 y). The results are shown in the table below.

n	200
Positives	0
Negatives	200
Prevalence	0.0%
	Ratio
Lowest value	0.0
Highest value	0.1
Mean value	0.0
Std deviation	0.01

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.

Michael Locke
Signature

Dir. Requlatory Affairs

12 June 2013 Date

{25}------------------------------------------------

Image /page/25/Picture/1 description: The image shows a close-up of a series of dark, rounded shapes arranged in a row. The shapes are evenly spaced and appear to be three-dimensional. The background is light, creating a strong contrast with the dark shapes. The shapes are positioned between two dark horizontal lines.

ATTACHMENT 1

PREMARKET NOTIFICATION 210(K) SAFETY AND EFFECTIVENESS SUMMARY (as required by 21 CFR § 807.92)

510(k)Number: A. K123261
Purpose for Submission: B. New device
C. Measurand:
Anti-Centromeres autoantibodies D. Type of Test:
- Qualitative enzyme immunoassay
E. Applicant:

EUROIMMUN US INC.
F. Proprietary and Established Names: EUROIMMUN Anti-Centromeres ELISA (IgG)
Regulatory Information: ઉ.
- Regulation: 1.
  - 21 CFR 866.5110 Antinuclear antibody immunological test system
- 1. Classification: Class II
- 1. Product code:
- LJM 4.
- Panel:
- Immunology Intended Use:
- 1. Intended use(s):

1. Indication(s) for use: Same as intended use.
1. Special conditions for the use statement(s): For prescription use only.
Special instrument requirements: 4.

Microwell plate reader capable of measuring OD at 450nm for dual wavelength readings.

l. Device Description:
The EUROIMMUN Anti-Centromeres ELISA (IgG) consists of a microwell ELISA plate coated with Centromeres antigen, calibrator, positive control, peroxidase-labelled anti-human IgG conjugate, sample buffer, wash buffer concentrate, TMB chromogen/substrate solution and stop solution.
J. Substantial Equivalence Information:
- Predicate device name (s): 1. Inova Quanta Lite Centromeres ELISA
- 1. Predicate 510(k) number(s): KOO3AEa

{26}------------------------------------------------

Image /page/26/Picture/1 description: The image shows a pattern of alternating black and white shapes. There are four black oval shapes that are evenly spaced. The black ovals are sandwiched between two black horizontal lines.

3. Comparison with predicate:

Similarities
Item	New Device	Predicate Device
Intended use	Detection of IgG antibodies to Centromeres	Same
Technology	ELISA	Same
Assay platform	96-well microtiter plates	Same
Calibration	Relative units	Same
Conjugate	Anti-human IgG labeled with horseradish peroxidase	Same
Substrate	TMB	Same
Reagent preparation	All reagents, calibrator and controls are ready to use,except for the wash buffer.	Same
Procedure	Sample incubation with micro-well antigen coated plate,followed by a wash step, incubation with an anti-humanIgG enzyme conjugate; wash step, incubation withsubstrate; then the addition of a stop solution andreading at 450nm.	Same
Differences
Item	New Device	Predicate Device
Assay format	Qualitative	"Semi-quantitative" (using low positive control)
Antigen	1:201	Recombinant CENP-A and CENP-B
Sample dilution	1 calibrator	1:101
Calibrators andcontrols	10x concentrate	3 controls: 1 high positive, 1 low positive (used forcalculation of results), 1 negative
Wash buffer	0.5 M sulphuric acid	40x concentrate
Stop solution	Serum or plasma (EDTA, Li-heparin, Citrate)	0.344 M sulphuric acid
Sample types	Ratio	Serum
Reported results	Ratio 1.0	Units
Cut off level	Qualitative	20 Units

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

Guidance for Industry and FDA Staff: Recommendations for Antibody (ANA) Test System Premarket (510(k)) Submissions (January 22, 2009)

L. Test Principle:

Patient samples are diluted 1:201 in sample buffer. 100 ul 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 µl 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 aqain washed 3 times with 300 ul of wash buffer to remove any unbound enzyme coniugate and 100 ul of 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. Performance Characteristics (where applicable):

1. Analytical performance:
- a. Precision/Reproducibility:

The reproducibility of the test was investigated using sera with different concentrations. Intra-assay reproducibility is based on 20 determinations and inter-assay reproducibility on 40 determinations performed in 10 different runs on 5 days with 2 runs per day, each run performed according to the package insert. The following results were obtained:

{27}------------------------------------------------

Image /page/27/Picture/0 description: The image shows a black and white illustration of a ladder. The ladder has two long, horizontal rails that run parallel to each other. There are four rungs connecting the two rails. The rungs are evenly spaced and have a rounded shape.

Intra-assay reproducibility		Anti-Centromeres ELISA (IgG)Ratio
n = 20		Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6
Mean value (x):	4.5	2.9	1.5	1.2	0.6	0.4
Range of values:	4.4 - 4.6	2.7 - 2.9	1.5 - 1.6	1.1 - 1.2	0.6 - 0.6	0.3 - 0.4
Expected result:.	positive	positive	positive	positive	negative	negative
% positive:	100%	100%	100%	100%	0%	0%
% negative:	0%	0%	0%	0%	100%	100%

Inter-assay reproducibility

n = 10 x 4	Anti-Centromeres ELISA (IgG)Ratio
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6
Mean value (x):	4.7	3.0	1.5	1.2	0.7	0.4
Range of values:	4.1 - 5.0	2.8 - 3.3	1.4 - 1.6	1.1 - 1.3	0.6 - 0.7	0.3 - 0.4
Expected result:	positive	positive	positive	positive	negative	negative
% positive:	100%	100%	100%	100%	0%	0%
% negative:	0%	0%	0%	0%	100%	100%

Lot to lot reproducibility

	Anti-Centromeres ELISA (IgG)
	Ratio
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6
n	6*	6*	6*	6*	6*	6*
Mean value (x):	4.1	4.9	3.9	0.3	0.9	1.1
Range of values:	3.8 - 4.5	4.6 - 5.3	3.7 - 4.2	0.3 - 0.3	0.8 - 0.9	1.1 - 1.2
Expected result:	positive	positive	positive	negative	negative	positive
% positive:	100%	100%	100%	0%	0%	100%
% negative:	0%	0%	0%	100%	100%	0%

	Anti-Centromeres ELISA (IgG)Ratio
	Sample 7	Sample 8	Sample 9	Sample 10
n	8**	10**	11**	10**
Mean value (x):	2.8	6.2	6.8	8.4
Range of values:	2.2 - 3.3	5.5 - 7.0	4.9 - 8.3	7.8 - 9.1
Expected result:	positive	positive	positive	positive
% positive:	100%	100%	100%	100%
% negative:	0%	0%	0%	0%

*3 lots x 2 runs ** n lots x 1 run

Linearity/assay reportable range: b. Not applicable.
High dose Hook effect C.

Potential for a high dose Hook effect is a phenomenon that is inherent with one step "sandwich" assay designs: Very high concentrations of antigen in the patient sample bind to all available sites saturating them - on both the antibody-solid phase and the antibody-labeled conjugate and thereby prevent the "sandwich" formation. Under these conditions, the measured level of analyte may be significantly lower than the actual level present in the sample. The two-step immunoassay design of the Anti-Centromeres ELISA (IgG) eliminates the adverse contribution of binding proteins. endogenous interfering substances and general matrix effects due to the extra wash step.

{28}------------------------------------------------

d. Traceability, Stability, Expected values (controls, calibrators or methods):

A recognized standard or reference material for anti-nuclear antibodies is not available. Results of this assay are given in ratios. The reactivity of the Anti-Centromeres ELISA was verified using the ANA reference panel of the CDC Centers for Disease Control and Prevention, Atlanta, USA.

Limit of detection: રે. Not applicable.
f. Analytical specificity:

Cross-reactivity: The quality of the antigen coated on the plates ensures a high specificity of the ELISA. Cross reactivity was investigated using a panel of 30 sera serologically positive for antibodies against rib.P-P, nRNP/Sm, Sm, SS-A, Scl-70, Jo-1, Rubella virus, Measles virus, Herpes simplex virus type 1 and Borrelia burgdorferi. All 30 sera were negative in the Anti-Centromeres ELISA (IgG), so no cross reactivity is expected.

Interference: To investigate the influence from hemoglobin, triglycerides and billrubin, 5 different sera at different anti-Centromeres concentrations (12 - 106 RU/ml) were spiked with potential interfering substances and were incubated with the test system according to the package insert. Interferences from rheumatoid factor were investigated similarly by spiking with a RF positive serum. The recovery in relation to the unspiked sample without interferent was calculated. The individual recovery was within the range of 91 - 111 %. No significant interference was observed for concentrations of up to 1000 mg/dl for hemoglobin, 2000 mg/dl for triglyceride, 40 mg/dl for billrubin and 500 IU/ml for rheumatoid factor.

ರು Assay cut-off:
Ratio 1.0

Comparison studies:

Method comparison with predicate device: a.
A comparison study was performed using 297 clinically characterized samples (144 systemic sclerosis, 2 CREST, 51 Sjögren's syndrome, 15 fibromyalgia, 35 RA, 30 borreliosis and 20 healthy), obtained from different sources from Europe and North America. The panel consisted of 61 men and 230 women. Age ranged from 2 to 87 vears with an average age of 50 years. Anti-centromeres antibodies are expected in systemic sclerosis. The other disease groups serve as control cohorts. The samples were tested with the EUROIMMUN Anti-Centromeres ELISA (IgG) and with the Inova Quanta Lite Centromere ELISA as the predicate device. All of the 5 discrepant samples were from control groups.

All samplesn = 297		Predicate ELISA
		positive	negative
EUROIMMUNAnti-Centromeres ELISA (IgG)	positive	73	0
	negative	5	219
Negative agreement	$219 / 219 = 100.0%$	95% C.I.: 98.3%	- 100.0%
Positive agreement	$73 / 78 = 93.6%$	95% C.I.: 85.7%	- 97.9%
Overall agreement	$292 / 297 = 98.3%$	95% C.I.: 96.1%	- 99.5%

b. Matrix comparison:

The usability of plasma was investigated using sample pairs each of serum and corresponding plasma (EDTA, Li-heparin, Citrate). Passing-Bablok regression was calculated for the comparison of serum to plasma. The regression equation is near the ideal correlation (intercept 0; slope 1.0) indicating equivalence of concentrations between serum and the corresponding plasma matrices. Coefficients of determination were found to be above 0.99 and %recovery compared toserum was in the range of 84 to 108 % (serum = 100 %).

	EDTA plasma	Li-heparin plasma	Citrate plasma
n	16	16	16
Regression equation(y = plasma, x = serum)	$y = -1.54 + 1.02 x$	$y = 0.02 + 0.92 x$	$y = -0.99 + 0.98$
95% C.I. of intercept	-3.85 – -0.06	-2.13 – 0.99	-2.48 – 0.69
95% C.I. of slope	0.98 – 1.08	0.89 – 0.96	0.94 – 1.01
Coefficient of determination R2	0.9958	0.9973	0.9978
Mean %recovery	98 %	93 %	96 %
Range of %recovery	84 – 108 %	83 – 101 %	88 – 107 %

{29}------------------------------------------------

3. Clinical studies:

Clinical studies were performed in cooperation with different sites. In total 877 clinically characterized samples (280 from systemic sclerosis patients and 597 from control groups) were investigated for anticentromeres antibodies (IgG). With the EUROIMMUN Anti-Centromeres ELISA (IgG) a prevalence of 19.0% (95% C.I.: 13.9 - 24.9%) was found in systemic sclerosis, a prevalence of 7.3% (95% C.I.: 3.0 -14.4%) was found in diffuse systemic sclerosis and a prevalence of 74.3% (95% C.I.: 65.3 - 82.1%) was found in limited systemic sclerosis with a specificity of 99.0% (95% C.I.; 97.8 - 99.6%). The results are shown in the table below. 95% C.I. are calculated by the exact method.

Clinical sensitivity: a.

No.	Panel	n	Anti-Centromeres ELISA (IgG)
			positive	%	95% C.I.
1	Systemic sclerosis	280	105	37.5%	31.8 - 43.5%
1a	Diffuse systemic sclerosis (from panel 1)	96	7	7.3%	3.0 - 14.4%
1b	Limited systemic sclerosis (from panel 1)	113	84	74.3%	65.3 - 82.1%

b. Clinical specificity:

No.	Panel	n.	negative	%	95% C.I.
2	Systemic lupus erythematosus	181	180	99.4%	97.0 - 100.0%
3	Polymyositis/dermatomyositis	26	26	100.0%	86.8 - 100.0%
4	Rheumatoid arthritis	164	163	99.4%	96.6 - 100.0%
5	Sjögren's syndrome	88	85	96.6%	90.4 - 99.3%
6	Mixed connective tissue diseases	45	44	97.8%	88.2 - 99.9%
7	Other autoimmune diseases*	63	63	100.0%	94.3 - 100.0%
8	Borreliosis	30	30	100.0%	88.4 - 100.0%
	Total	597	591	99.0%	97.8 - 99.6%

*from the following groups: AIH (n = 8), Grave's disease (n = 12), Hashimoto (n = 11), celiac disease (n = 11), Diabetes Type I (n = 12)

Other clinical supportive data (when a. and b. are not applicable): C.
- Not applicable.
1. Clinical cut-off:

See Assay cut-off.

ട്. Expected values/Reference range:
The levels of anti-centromeres antibodies (IgG) were analyzed in a panel of 200 samples from apparently healthy blood donors (120 men and 80 women with an average age of 40 y; age range: 19 - 68 y). The results are shown in the table below.

n	200
Positives	1
Negatives	199
Prevalence	0.5%
	Ratio
Lowest value	0.0
Highest value	3.0
Mean value	0.1
Std deviation	0.21

· 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.

Michael Locke
Signature

Dir. Regulatory Affairs

12 June 2013 Dote

{30}------------------------------------------------

Image /page/30/Figure/1 description: The image shows a pattern of alternating black and white shapes. The black shapes appear to be horizontal bars running across the top and bottom of the image. In between the bars are a series of black oval shapes, separated by white space. There are four black oval shapes in total.

ATTACHMENT I

PREMARKET NOTIFICATION 210(K) SAFETY AND EFFECTIVENESS SUMMARY (as required by 21 CFR § 807.92)

A. 510(k)Number:

K123261

Purpose for Submission: B. New device
C. Measurand:
- Anti-Jo-1 autoantibodies
D. Type of Test: Qualitative enzyme immunoassay
E. Applicant:
- EUROIMMUN US INC.
F. Proprietary and Established Names: EUROIMMUN Anti-Jo-1 ELISA (IgG)

ઉં. Requlatory Information:

1. Regulation:
- 21 CFR 866.5110 Antinuclear antibody immunological test system
1. Classification: Class II
1. Product code: LLL
ধ . Panel:

Immunology Intended Use:
Intended use(s): 1.

The EUROIMMUN Anti-Jo-1 ELISA (IqG) test kit is intended for the qualitative determination of IgG class autoantibodies against Jo-1 in human serum and plasma (EDTA, Li-heparin, Citrate). It is used as an aid in the diagnosis of polymyositis and dermatomyositis, in conjunction with other laboratory and clinical findings.

1. Indication(s) for use: Same as intended use.
1. Special conditions for the use statement(s): For prescription use only.

Special instrument requirements:

Microwell plate reader capable of measuring OD at 450nm for dual wavelength readings.

l. Device Description:

۔ ل۔ Substantial Equivalence Information:

Predicate device name (s): 1.
- Inova Quanta Lite Jo-1 ELISA
1. Predicate 510(k) number(s): Kazesez

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3. Comparison with predicate:

Similarities
Item	New Device	Predicate Device
Intended use	Detection of IgG antibodies to Jo-1	Same
Technology	ELISA	Same
Assay platform	96-well microtiter plates	Same
Calibration	Relative units	Same
Antigen	Purified Jo-1 antigen	Same
Conjugate	Anti-human IgG labeled with horseradish peroxidase	Same
Substrate	TMB	Same
Reagent preparation	All reagents, calibrator and controls are ready to use,except for the wash buffer.	Same
Procedure	Sample incubation with micro-well antigen coated plate,followed by a wash step, incubation with an anti-humanIgG enzyme conjugate; wash step, incubation withsubstrate; then the addition of a stop solution andreading at 450nm.	Same
Differences
Item	New Device	Predicate Device
Assay format	Qualitative	"Semi-quantitative" (using low positive control)
Sample dilution	1:201	1:101
Calibrators and	1 calibrator	3 controls: 1 high positive, 1 low positive (used for
controls	2 controls: 1 positive, 1 negative	calculation of results), 1 negative
Wash buffer	10x concentrate	40x concentrate
Stop solution	0.5 M sulphuric acid	0.344 M sulphuric acid
Sample types	Serum or plasma (EDTA, Li-heparin, Citrate)	Serum
Reported results	Ratio	Units
Cut off level	Ratio 1.0	20 Units

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

Guidance for Industry and FDA Staff: Recommendations for Antibody (ANA) Test System Premarket (510(k)) Submissions (January 22, 2009)

L. Test Principle:

Patient samples are diluted 1:201 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 coniugate 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 ul of wash buffer to remove any unbound-enzyme conjugate and 100 µl of 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. Performance Characteristics (where applicable):

Analytical performance: 1.

Precision/Reproducibility: a.
The reproducibility of the test was investigated using sera with different concentrations. Intra-assay reproducibility is based on 20 determinations and inter-assay reproducibility on 40 determinations performed in 10 different runs on 5 days with 2 runs per day, each run performed according to the package insert. The following results were obtained:

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Image /page/32/Picture/0 description: The image shows a black and white pattern. The pattern consists of four black shapes that are separated by white spaces. The black shapes are connected by two horizontal black lines, one at the top and one at the bottom. The shapes are symmetrical and have a curved appearance.

Intra-assay reproducibility

	Anti-Jo-1 ELISA ELISA (IgG)Ratio
n = 20
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6
Mean value (x):	6.4	4.3	1.8	1.0	0.8	0.4
Range of values:	6.1 - 6.8	4.1 - 4.5	1.6 - 1.9	1.0 - 1.1	0.7 - 0.8	0.3 - 0.4
Expected result:	positive	positive	positive	positive	negative	negative
% positive:	100%	100%	100%	100%	0%	0%
% negative:	0%	0%	0%	0%	100%	100%

Inter-assay reproducibility

n = 10 x 4	Anti-Jo-1 ELISA (IgG)Ratio
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6
Mean value (x):	6.0	4.2	2.2	1.1	0.8	0.4
Range of values:	5.6 - 6.6	3.7 - 4.6	2.0 - 2.3	1.0 - 1.2	0.7 – 0.8	0.3 - 0.4
Expected result:	positive	positive	positive	positive	negative	negative
% positive:	100%	100%	100%	100%	0%	0%
% negative:	0%	0%	0%	0%	100%	100%

Lot to lot reproducibility

	Anti-Jo-1 ELISA (IgG) Ratio								Anti-Jo-1 ELISA (IgG)Ratio
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6		Sample 7	Sample 8	Sample 9	Sample 10
n	6*	6*	6*	6*	6*	6*	n	10**	11**	11**	11**
Mean value (x):	5.1	3.9	2.0	0.3	0.9	1.1	Mean value (x):	2.6	4.0	7.4	8.2
Range of values:	4.6 - 5.4	3.5 - 4.4	1.9 - 2.2	0.3 - 0.4	0.9 - 0.9	1.1 - 1.2	Range of values:	1.9 - 3.1	3.2 – 4.6	6.4 - 8.6	7.7 - 8.9
Expected result:	positive	positive	positive	negative	negative	positive	Expected result:	positive	positive	positive	positive
% positive:	100%	100%	100%	0%	0%	100%	% positive:	100%	100%	100%	100%
% negative:	0%	0%	0%	100%	100%	0%	% negative:	0%	0%	0%	0%

*3 lots x 2 runs ** n lots x 1 run

Linearity/assay reportable range: b. Not applicable.
High dose Hook effect C.

Potential for a high dose Hook effect is a phenomenon that is inherent with one step "sandwich" assay designs: Very high concentrations of antigen in the patient sample bind to all available sites saturating them - on both the antibody-solid phase and the antibody-labeled conjugate and thereby prevent the "sandwich" formation. Under these conditions, the measured level of analyte may be significantly lower than the actual level present in the sample. The two-step immunoassay design of the Anti-Jo-1 ELISA (IgG) eliminates the adverse contribution of binding proteins, endogenous interfering substances and general matrix effects due to the extra wash step.

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Image /page/33/Picture/1 description: The image shows a black and white illustration of a decorative border. The border consists of a series of rounded shapes, resembling beads or small bulbs, positioned between two horizontal lines. The shapes are evenly spaced and create a repeating pattern along the length of the border.

d. Traceability, Stability, Expected values (controls, calibrators or methods):

A recognized standard or reference material for anti-nuclear antibodies is not available. Results of this assay are given in ratios. The reactivity of the Anti-Jo-1 ELISA was verified using the ANA reference panel of the CDC Centers for Disease Control and Prevention, Atlanta, USA.

Limit of detection: e.
Not applicable. f. Analytical specificity:

Cross-reactivity: The quality of the antigen coated on the plates ensures a high specificity of the ELISA. Cross reactivity was investigated using a panel of 30 sera serologically positive for antibodies against rib.P-P, nRNP/Sm, Sm, SS-A, SS-B, Scl-70, centromeres, Rubella virus, Measles virus, Herpes simplex virus type 1 and Borrelia burgdorferi. All 30 sera were negative in the Anti-Jo-1 ELISA (IgG), so no cross reactivity is expected.

Interference: To investigate the influence from hemoglobin, triglycerides and bilirubin, 5 different sera at different anti-Jo-1 concentrations (15 - 106 RU/ml) were spiked with potential interfering substances and were incubated with the test system according to the package insert. Interferences from rheumatoid factor were investigated similarly by spiking with a RF positive serum. The recovery in relation to the unspiked sample without interferent was calculated. The individual recovery was within the range of 91 - 122 %. No significant interference was observed for concentrations of up to 1000 mg/dl for hemoglobin, 2000 mg/dl for triglyceride, 40 mg/dl for bilirubin and 500 lU/ml for rheumatoid factor.

Assay cut-off: ರು
Ratio 1.0

2. Comparison studies:

Method comparison with predicate device: a.
A comparison study was performed using 297 clinically characterized samples (143 myositis, 3 PM/SSc, 51 Sjögren's syndrome, 15 fibromyalgia, 35 RA, 30 borreliosis and 20 healthy), obtained from different sources from Europe and North America. The panel consisted of 75 men and 221 women. Age ranged from 14 to 84 years with an average age of 52 years. Anti-Jo-1 antibodies are expected in myositis. The other disease groups serve as control cohorts. The samples were tested with the EUROIMMUN Anti-Jo-1 ELISA (IgG) and with the Inova Quanta Lite Jo-1 ELISA as the predicate device. Both discrepant samples were from myositis patients.

All samplesn = 297		Predicate ELISA
		positive	negative
EUROIMMUNAnti-Jo-1 ELISA (IgG)	positive	64	1
	negative	1	231
Negative agreement		231 / 232 = 99.6%	95% C.I.: 97.6% - 100.0%
Positive agreement		64 / 65 = 98.5%	95% C.I.: 91.7% - 100.0%
Overall agreement		295 / 297 = 99.3%	95% C.I.: 97.6% - 99.9%

b. Matrix comparison:
The usability of plasma was investigated using sample pairs each of serum and corresponding plasma (EDTA, Li-heparin, Citrate). Passing-Bablok regression was calculated for the comparison of serum to plasma. The regression equation is near the ideal correlation (intercept 0; slope 1.0) indicating equivalence of concentrations between serum and the corresponding plasma matrices. Coefficients of determination were found to be above 0.98 and %recovery compared toserum was in the range of 85 to 117 % (serum = 100 %).

	EDTA plasma	Li-heparin plasma	Citrate plasma
n	15	15	15
Concentration range (serum)	ratio 0.3 - 9.2	ratio 0.3 - 9.2	ratio 0.3 - 9.2
Concentration range (plasma)	ratio 0.3 - 8.7	ratio 0.4 - 8.5	ratio 0.3 - 8.9
Regression equation(y = plasma, x = serum)	y = -0.09 + 0.99 x	y = 0.09 + 0.93 x	y = 0.73 + 0.99
95% C.I. of intercept	-3.77 - 1.83	-0.69 - 1.94	-0.49 - 2.66
95% C.I. of slope	0.93 - 1.10	0.89 - 0.99	0.96 - 1.01
Coefficient of determination R2	0.9856	0.9956	0.9982
Mean %recovery	101 %	95 %	103 %
Range of %recovery	85 - 117 %	87 - 113 %	94 - 114 %

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3. Clinical studies:

Clinical studies were performed in cooperation with different sites. In total 876 clinically characterized samples (177 from myositis patients and 699 from control groups) were investigated for anti-Jo-1 antibodies (IgG). With the EUROIMMUN Anti-Jo-1 ELISA (IgG) a prevalence of 18.6% (95% C.I.: 13.2 – 25.2%) was found in myositis with a specificity of 99.6% (95% C.I.: 98.8 – 99.9%). The results are shown in the table below. 95% C.I. are calculated by the exact method. "

a. Clinical sensitivity:

NO.	Panel		FLISA (IaG)Anti-Jo-1
			positive		95% C.I.
	------------------------------------------------------------------------------------------------------------------------------------------------------------------------------Polymvositis/dermatomyositis	1 79 97		18.6%	25.2%1 - 25.6 .
No.	Panel	n	Anti-Jo-1 ELISA (IgG)
			negative	%	95% C.I.
2	Systemic lupus erythematosus	213	211	99.1%	96.6 - 99.9%
3	Rheumatoid arthritis	164	163	99.4%	96.6 - 100.0%
4	Systemic sclerosis	81	81	100.0%	95.5 - 100.0%
5	Sjögren's syndrome	88	88	100.0%	95.9 - 100.0%
6	Mixed connective tissue diseases	45	45	100.0%	92.1 - 100.0%
7	Fibromyalgia	15	15	100.0%	78.2 - 100.0%
8	Other autoimmune diseases	63	63	100.0%	94.3 - 100.0%
9	Borreliosis	30	30	100.0%	88.4 - 100.0%
	Total	699	696	99.6%	98.8 - 99.9%

b. Clinical specificity:

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

Not applicable.
ব : Clinical cut-off:

See Assay cut-off.

1. Expected values/Reference range:
  The levels of antibodies (IgG) were analyzed in a panel of 200 samples from apparently healthy blood donors (120 men and 80 women with an average age of 40 y; age range: 19 - 68 y). The results are shown in the table below.

n	200
Positives	0
Negatives	200
Prevalence	0.0%
	Ratio
Lowest value	0.0
Highest value	0.2
Mean value	0.1
Std deviation	0.03

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.

Michael Locke
Signature

Dir. Regulatory Affairs Title

12 June 2013 Date

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ATTACHMENT I

PREMARKET NOTIFICATION 200(K) SAFETY AND EFFECTIVENESS SUMMARY (as required by 21 CFR § 807.92)

A. 510(k)Number:
- K123261
B. Purpose for Submission: · New device
C. Measurand:
- Anti-ribosomal P-proteins autoantibodies
D. Type of Test:
- Qualitative enzyme immunoassay
E. Applicant: EUROIMMUN US INC.
F. Proprietary and Established Names: EUROIMMUN Anti-ribosomal P-proteins ELISA (IgG)

G. Requlatory Information:

1. Regulation:
- 21 CFR 866.5110 Antinuclear antibody immunological test system
1. Classification: Class II
1. Product code: MQA -
র . Panel:

Immunology
Intended Use:
1. Intended use(s):

1. Indication(s) for use: Same as intended use.
1. Special conditions for the use statement(s): For prescription use only.

র্ব . Special instrument requirements:

Microwell plate reader capable of measuring OD at 450nm for dual wavelength readings. Device Description:

J. Substantial Equivalence Information:

Predicate device name (s): 1.
- Inova Quanta Lite Ribosomal P ELISA
1. Predicate 510(k) number(s): K981237

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Image /page/36/Picture/1 description: The image shows the number 3 followed by the words "Comparison with predicate:". The text is black and the background is white. The text is left-aligned.

SimilaritiesItem	New Device	Predicate Device
Intended use	Detection of IgG antibodies to ribosomal P-proteins	Same
Technology	ELISA	Same
Assay platform	96-well microtiter plates	Same
Calibration	Relative units	Same
Conjugate	Anti-human IgG labeled with horseradish peroxidase	Same
Substrate	TMB	Same
Reagent preparation	All reagents, calibrator and controls are ready to use,except for the wash buffer.	Same
Procedure	Sample incubation with micro-well antigen coated plate,followed by a wash step, incubation with an anti-humanIgG enzyme conjugate; wash step, incubation withsubstrate; then the addition of a stop solution andreading at 450nm.	Same
Differences
Item	New Device	Predicate Device
Assay format	Qualitative	"Semi-quantitative" (using low positive control)
Antigen	Purified ribosomal P antigen	Synthetic ribosomal P antigen
Sample dilution	1:201	1:101
Calibrators andcontrols	1 calibrator2 controls: 1 positive, 1 negative	3 controls: 1 high positive, 1 low positive (used forcalculation of results), 1 negative
Wash buffer	10x concentrate	40x concentrate
Stop solution	0.5 M sulphuric acid	0.344 M sulphuric acid
Sample types	Serum or plasma (EDTA, Li-heparin, Citrate)	Serum
Reported results	Ratio	Units
Cut off level	Ratio 1.0	20 Units

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

Guidance for Industry and FDA Staff: Recommendations for Antibody (ANA) Test System Premarket (510(k)) Submissions (January 22, 2009)

L. Test Principle:

Patient samples are diluted 1:201 in sample buffer, 100 pl 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 microtier 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 ul of wash buffer to remove any unbound enzyme conjuqate and 100 ul of the chromogen substrate is added. The strips are incubated for 15 minutes at room temperature and 100 ul stop solution is added. The microtier 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.

Performance Characteristics (where applicable): M.

Analytical performance: 1.
- Precision/Reproducibility: a.

The reproducibility of the test was investigated using sera with different concentrations. Intra-assay reproducibility is based on 20 determinations and inter-assay reproducibility on 40 determinations performed in 10 different runs on 5 days with 2 runs per day, each run performed according to the package insert. The following results were obtained:

{37}------------------------------------------------

Image /page/37/Picture/0 description: The image shows a black and white illustration of a decorative border. The border consists of a series of oval or rounded shapes, evenly spaced and connected by horizontal lines above and below. The shapes are solid black, creating a stark contrast against the white background.

Intra-assay reproducibility

	Anti-ribosomal P-proteins ELISA (IgG)Ratio
n = 20
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6
Mean value (x):	5.5	3.6	1.9	1.0	0.6	0.4
Range of values:	5.1 - 6.0	3.2 - 4.1	1.8 - 2.1	1.0 - 1.1	0.6 - 0.7	0.3 - 0.5
Expected result:	positive	positive	positive	positive	negative	negative
% positive:	100%	100%	100%	100%	0%	0%
% negative:	0%	0%	0%	0%	100%	100%

Inter-assay reproducibility

n = 10 x 4	Anti-ribosomal P-proteins ELISA (IgG)Ratio
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6
Mean value (x):	5.7	3.8	1.9	1.1	0.7	0.4
Range of values:	5.3 - 6.1	3.5 - 4.1	1.7 - 2.2	1.0 - 1.3	0.6 - 1.0	0.3 - 0.4
Expected result:	positive	positive	positive	positive	negative	negative
% positive:	100%	100%	100%	100%	2.5%	0%
% negative:	0%	0%	0%	0%	97.5%	100%

Lot to lot reproducibility

	Anti-ribosomal P-proteins ELISA (IgG)Ratio
	Sample 1	Sample 2	Sample 3	Sample 4	Sample 5	Sample 6
n	6*	6*	6*	6*	6*	6*
Mean value (x):	3.2	4.5	2.2	0.3	0.9	1.1
Range of values:	2.8 – 3.6	4.2 - 4.8	2.0 - 2.4	0.3-0.4	0.8 - 0.9	1.0 - 1.2
Expected result:	positive	positive	positive	negative	negative	positive
% positive:	100%	100%	100%	0%	0%	100%
% negative:	0%	0%	0%	100%	100%	0%

	Anti-ribosomal P-proteins ELISA (IgG)Ratio
	Sample 7	Sample 8	Sample 9
n	11**	11**	11**
Mean value (x):	4.8	5.2	7.6
Range of values:	3.9 - 5.3	4.4 - 6.2	6.5 – 8.5
Expected result:	positive	positive	positive
% positive:	100%	100%	100%
% negative:	0%	0%	0%

*3 lots x 2 runs ** n lots x 1 run

Linearity/assay reportable range: b. Not applicable.
High dose Hook effect C.

Potential for a high dose Hook effect is a phenomenon that is inherent with one step "sandwich" assay designs: Very high concentrations of antigen in the patient sample bind to all available sites saturating them - on both the antibody-solid phase and the antibody-labeled conjugate and thereby prevent the "sandwich" formation. Under these conditions, the measured level of analyte may be significantly lower than the actual level present in the sample. The two-step immunoassay design of the Anti-ribosomal P-proteins ELISA (IgG) eliminates the adverse contribution of binding proteins, endogenous interfering substances and general matrix effects due to the extra wash step.

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Image /page/38/Picture/1 description: The image shows a black and white pattern. The pattern consists of a series of black rectangles connected by black, rounded shapes. The rectangles are arranged horizontally, and the rounded shapes are positioned between them, creating a repeating sequence.

Traceability. Stability, Expected values (controls, calibrators or methods): · d.

A recognized standard or reference material for anti-nuclear antibodies is not available. Results of this assay are given in ratios. The reactivity of the Anti-ribosomal P-proteins ELISA was verified using the ANA reference panel of the CDC Centers for Disease Control and Prevention, Atlanta, USA.

Limit of detection: ୧.
- Not applicable.
r. Analytical specificity:

Cross-reactivity: The quality of the antigen coated on the plates ensures a high specificity of the ELISA. Cross reactivity was investigated using a panel of 30 sera serologically positive for antibodies against nRNP/Sm, Sm, SS-A, Scl-70, Jo-1, centromeres, Rubella virus, Measles virus, Herpes simplex virus type 1 and Borrelia burgdorferi. All 30 sera were negative in the Anti-ribosomal Pproteins ELISA (IgG), so no cross reactivity is expected.

Interference: To investigate the influence from hemoglobin, triglycerides and bilirubin, 5 different sera at different anti-ribosomal P-proteins concentrations (13 - 104 RU/ml) were spiked with potential interfering substances and were incubated with the test system according to the package insert. Interferences from rheumatoid factor were investigated similarly by spiking with a RF positive serum. The recovery in relation to the unspiked sample without interferent was calculated. The individual recovery was within the range of 84 - 115 %. No significant interference was observed for concentrations of up to 1000 mg/dl for hemoglobin, 2000 mg/dl for triglyceride, 40 mg/dl for bilirubin and 500 IU/ml for rheumatoid factor.

Assay cut-off: ರು
Ratio 1.0
1. Comparison studies:
- Method comparison with predicate device: a.

A comparison study was performed using 243 clinically characterized samples (90 SLE, 1 SLE/SS, 1 CLE, 51 Siögren's syndrome, 15 fibromyalgia, 35 RA, 30 borreliosis and 20 healthy), obtained from different sources from Europe and North America. The panel consisted of 49 men and 187 women. Age ranged from 14 to 83 years with an average age of 47 years. Anti-ribosomal P-proteins antibodies are expected in SLE. The other disease groups serve as control cohorts. The samples were tested with the EUROIMMUN Anti-ribosomal P-proteins ELISA (IgG) and with the Inova Quanta Lite Ribosomal P ELISA as the predicate device. All of the 10 discrepant samples were from patients with SLE, SLE/SS, and CLE.

All samplesn = 243	Predicate ELISA
	positive	negative
EUROIMMUNAnti-ribosomal P-proteinsELISA (IgG)	28	10
negative	0	205
Negative agreementPositive agreementOverall agreement	$205 / 215 = 95.3%$$28 / 28 = 100.0%$$233 / 243 = 95.9%$	95% C.I.: 91.6% - 97.7%95% C.I.: 87.7% - 100.0%95% C.I.: 92.6% - 98.0%

b. Matrix comparison:
The usability of plasma was investigated using sample pairs each of serum and corresponding plasma (EDTA, Li-heparin, Citrate). Passing-Bablok regression was calculated for the comparison of serum to plasma. The regression equation is near the ideal correlation (intercept 0; slope 1.0) indicating equivalence of concentrations between serum and the corresponding plasma matrices. Coefficients of determination were found to be above 0.99 and %recovery compared toserum was in the range of 79 to 107 % (serum = 100 %).

	EDTA plasma	Li-heparin plasma	Citrate plasma
n	16	16	16
Regression equation(y = plasma, x = serum)	$y = -0.98 + 1.02 x$	$y = 0.18 + 0.98 x$	$y = -1.79 + 0.99 x$
95% C.I. of intercept	-5.78 - 0.56	-2.42 - 3.26	-5.55 - 0.25
95% C.I. of slope	0.99 - 1.06	0.92 - 1.03	0.97 - 1.04
Coefficient of determination R2	0.9966	0.9939	0.9959
Mean %recovery	97 %	98 %	95 %
Range of %recovery	79 - 107 %	80 - 107 %	85 - 107 %

{39}------------------------------------------------

3. Clinical studies:

Clinical studies were performed in cooperation with different sites. In total 876 clinically characterized samples (376 from SLE patients and 500 from control groups) were investigated for anti-ribosomal Pproteins antibodies (IgG). With the EUROIMMUN Anti-ribosomal P-proteins ELISA (IgG) a prevalence of 5.3% (95% C.I.; 3.3 - 8.1%) was found in SLE with a specificity of 99.2% (95% C.I.: 98.0 - 99.8%). The results are shown in the table below. 95% C.I. are calculated by the exact method.

Clinical sensitivity: a.

No.	Panel	n	Anti-ribosomal P-proteins ELISA (IgG)
			positive	%	95% C.I.
1	Systemic lupus erythematosus	376	20	5.3%	3.3 - 8.1%
No.	Panel	n	Anti-ribosomal P-proteins ELISA (IgG)
			negative	%	95% C.I.
2	Polymyositis/dermatomyositis	151	149	98.7%	95.3 - 99.8%
3	Rheumatoid arthritis	90	90	100.0%	96.0 - 100.0%
4	Systemic sclerosis	66	66	100.0%	94.6 - 100.0%
5	Sjögren's syndrome	55	54	98.2%	90.3 - 100.0%
6	Mixed connective tissue diseases	45	44	97.8%	88.2 - 99.9%
7	Other autoimmune diseases*	63	63	100.0%	94.3 - 100.0%
8	Borreliosis	30	30	100.0%	88.4 - 100.0%
	Total	500	496	99.2%	98.0 - 99.8%

b. Clinical specificity:

from the following groups: AIH (n = 8), Grave's disease (n = 12), Hashimoto (n = 11), celiac disease (n = 11), Diabetes Type I (n = 12)

Other clinical supportive data (when a. and b. are not applicable): C. Not applicable.
Clinical cut-off: See Assay cut-off.
1. Expected values/Reference range:

The levels of anti-ribosomal P-proteins antibodies (IgG) were analyzed in a panel of 150 samples from apparently healthy blood donors (79 men and 71 women with an average age of 38 y; age range: 18 – 67 y). The results are shown in the table below.

n	150
Positives	0
Negatives	150
Prevalence	0.0%
	Ratio
Lowest value	0.0
Highest value	0.7
Mean value	0.1
Std deviation	0.08

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.

Michael Locke
Signature

Dir. Regulatory Affairs Title

12 June 2013
Date

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Image /page/40/Picture/0 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a stylized eagle with three stripes forming its wing and body. The eagle is positioned to the right of the text, which is arranged in a circular pattern around the logo. The text reads "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA".

DEPARTMENT OF HEALTH & HUMAN SERVICES

Public Health Service

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

June 12, 2013

THE BINDING SITE GROUP LTD C/O MS. KATHRYN KOHL MANAGING DIRECTOR 1100 THE AMERICAN ROAD MORRIS PLAINS, NJ 07950

Re: K123261

Trade/Device Name: Euroimmun Anti-nRNP/Sm ELISA (IgG) Euroimmun Anti-Sm ELISA (IgG) Euroimmun Anti-SS-A ELISA (IgG) Euroimmun Anti-SS-B ELISA (IgG) Euroimmun Anti-Scl-70 ELISA (IgG) Euroimmun Anti-Centromeres ELISA (IgG) Euroimmun Anti-Jo-1 ELISA (IgG) Euroimmun Anti-Ribosomal P-Proteins ELISA (IgG) Regulation Number: 21 CFR 866.5110 Regulation Name: Antinuclear Antibody Immunological Test System Regulatory Class: II Product Code: LKO, LKP, LLL, LJM, MQA Dated: May 16, 2013 Received: May 17, 2013

Dear Ms. Kohl:

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.

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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 (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 regulations (21 CFR Parts 801 and 809), please contact 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/Industrv/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.fdai.gov/MedicalDevices/Safetv/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 Center for Devices and Radiological Health

Enclosure

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510(k) Number (if known): K123261

Device Name: Anti-ribosomal P-proteins ELISA (IgG)

Indications For Use:

Prescription Use × -(Part-21-CFR-801-Subpart-D)- AND/OR

Over-The-Counter Use -(21-CFR-807-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)

Maria M. Chan -S

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

510(k): k 123261

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510(k) Number (if known): K123261

Device Name: Anti-nRNP/Sm ELISA (IqG)

Indications For Use:

Prescription Use × (Part 21 CFR 801 Subpart D) AND/OR

Over-The-Counter Use (21 CFR 807 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)

Maria M. Chan -S

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

510(k):

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510(k) Number (if known): K123261

Device Name: Anti-Sm ELISA (IgG)

Indications For Use:

The EUROIMMUN Anti-Sm ELISA (IgG) test kit is intended for the gualitative determination of IgG class autoantibodies against Sm in human serum and plasma (EDTA, Li-heparin, Citrate). It is used as an aid in the diagnosis of systemic lupus erythematosus, in conjunction with other laboratory and clinical findings.

Prescription Use × (Part 21 CFR 801 Subpart D) AND/OR

Over-The-Counter Use (21 CFR 807 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)

Maria M. Chan -S

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

{45}------------------------------------------------

510(k) Number (if known): K123261

Device Name: Anti-SS-A ELISA (IgG)

Indications For Use:

Prescription Use × (Part 21 CFR 801 Subpart D) Over-The-Counter Use

(21 CFR 807 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): K123261

{46}------------------------------------------------

510(k) Number (if known): K123261

Device Name: Anti-SS-B ELISA (IgG)

Indications For Use:

Prescription Use × (Part 21 CFR 801 Subpart D) Over-The-Counter Use (21 CFR 807 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

{47}------------------------------------------------

510(k) Number (if known): K123261

Device Name: Anti-Scl-70 ELISA (IgG)

Indications For Use:

The EUROIMMUN Anti-Scl-70 ELISA (IgG) test kit is intended for the qualitative determination of IgG class autoantibodies against Scl-70 in human serum and plasma (EDTA, Li-heparin, Citrate). It is used as an aid in the diagnosis of progressive systemic sclerosis, in conjunction with other laboratory and clinical findings.

Prescription Use × (Part 21 CFR 801 Subpart D) AND/OR

Over-The-Counter Use (21 CFR 807 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)

Maria M. Chan -S

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

{48}------------------------------------------------

510(k) Number (if known): K123261

Device Name: Anti-Centromeres ELISA (IgG)

Indications For Use:

The EUROIMMUN Anti-Centromeres ELISA (IgG) test kit is intended for the qualitative determination of IgG class autoantibodies against centromeres in human serum and plasma (EDTA, Li-heparin, Citrate). It is used as an aid in the diagnosis of limited form of progressive systemic sclerosis (CREST syndrome), in conjunction with other laboratory and clinical findings.

Prescription Use X (Part 21 CFR 801 Subpart D) Over-The-Counter Use (21 CFR 807 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

{49}------------------------------------------------

510(k) Number (if known): K123261

Device Name: Anti-Jo-1 ELISA (IgG)

Indications For Use:

Prescription Use × (Part 21 CFR 801 Subpart D) AND/OR

Over-The-Counter Use (21 CFR 807 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)

Maria M. Chan -S

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

510(k): K123261

Regulation Number and Section

§ 866.5100 Antinuclear antibody immunological test system.

(a)
Identification. An antinuclear antibody immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the autoimmune antibodies in serum, other body fluids, and tissues that react with cellular nuclear constituents (molecules present in the nucleus of a cell, such as ribonucleic acid, deoxyribonucleic acid, or nuclear proteins). The measurements aid in the diagnosis of systemic lupus erythematosus (a multisystem autoimmune disease in which antibodies attack the victim's own tissues), hepatitis (a liver disease), rheumatoid arthritis, Sjögren's syndrome (arthritis with inflammation of the eye, eyelid, and salivary glands), and systemic sclerosis (chronic hardening and shrinking of many body tissues).(b)
Classification. Class II (performance standards).