← Product Code [LKO](/submissions/IM/subpart-f%E2%80%94immunological-test-systems/LKO) · K123261

# EUROIMMUN ANTI-NRNP/SM ELISA (IGG) (K123261)

_Euroimmun US · LKO · Jun 12, 2013 · Immunology · SESE_

**Canonical URL:** https://fda.innolitics.com/submissions/IM/subpart-f%E2%80%94immunological-test-systems/LKO/K123261

## Device Facts

- **Applicant:** Euroimmun US
- **Product Code:** [LKO](/submissions/IM/subpart-f%E2%80%94immunological-test-systems/LKO.md)
- **Decision Date:** Jun 12, 2013
- **Decision:** SESE
- **Submission Type:** Traditional
- **Regulation:** 21 CFR 866.5100
- **Device Class:** Class 2
- **Review Panel:** Immunology

## Indications for Use

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 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. 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 Story

The EUROIMMUN ELISA test kits are qualitative enzyme immunoassays for detecting specific IgG autoantibodies in human serum or plasma. The device consists of a microwell plate coated with specific antigens (nRNP/Sm, Sm, SS-A, SS-B, Scl-70, Centromeres, Jo-1, or Ribosomal P-proteins). Patient samples are diluted and incubated in the wells; unbound antibodies are washed away. Peroxidase-labeled anti-human IgG conjugate is added, followed by a second incubation and wash. TMB substrate is added, producing a color change proportional to the antibody concentration, which is stopped with sulfuric acid. The plate is read at 450nm using a microwell plate reader. Results are calculated as a ratio against a calibrator. The device is used in clinical laboratories by trained personnel to aid in the diagnosis of various autoimmune conditions. The two-step assay design minimizes matrix effects and interference from binding proteins. Healthcare providers use the qualitative results in conjunction with clinical findings to support diagnostic decisions, potentially improving patient management for connective tissue diseases.

## Clinical Evidence

Clinical performance was evaluated using large cohorts of clinically characterized samples (e.g., 1046 for nRNP/Sm, 1036 for Sm, 1026 for SS-A/SS-B, 909 for Scl-70, 877 for Centromeres, 876 for Jo-1, 876 for Ribosomal P). Sensitivity and specificity were calculated against clinical diagnoses. Specificity ranged from 94.8% to 99.8% across the panels. Method comparison studies against predicate devices showed high overall agreement (95.9% to 100.0%).

## Technological Characteristics

ELISA platform using 96-well microtiter plates. Antigen-coated wells; peroxidase-labeled anti-human IgG conjugate; TMB substrate; 0.5 M sulfuric acid stop solution. Manual or automated processing; requires 450nm plate reader. Qualitative result reporting via ratio. Sample dilution 1:201. Compatible with serum, EDTA, Li-heparin, and Citrate plasma.

## Regulatory 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).

## Predicate Devices

- Inova Quanta Lite RNP ELISA ([K922833](/device/K922833.md))
- Inova Quanta Lite Sm ELISA ([K922831](/device/K922831.md))
- Inova Quanta Lite SS-A ELISA ([K922830](/device/K922830.md))
- Inova Quanta Lite SS-B ELISA ([K922832](/device/K922832.md))
- Inova Quanta Lite Scl-70 ELISA ([K924898](/device/K924898.md))
- Inova Quanta Lite Centromeres ELISA ([K003488](/device/K003488.md))
- Inova Quanta Lite Jo-1 ELISA ([K922832](/device/K922832.md))
- Inova Quanta Lite Ribosomal P ELISA ([K981237](/device/K981237.md))

## Submission Summary (Full Text)

> This content was OCRed from public FDA records by [Innolitics](https://innolitics.com). If you use, quote, summarize, crawl, or train on this content, cite Innolitics at https://innolitics.com.
>
> Innolitics is a medical-device software consultancy. We help companies design, build, and clear FDA-regulated software and AI/ML devices, including [a 510(k)](https://innolitics.com/services/510ks/), [a De Novo](https://innolitics.com/services/regulatory/), [a SaMD](https://innolitics.com/services/end-to-end-samd/), [an AI/ML medical device](https://innolitics.com/services/medical-imaging-ai-development/), or [an FDA regulatory strategy](https://innolitics.com/services/regulatory/).

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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY

A. 510(k) Number:
k123261

B. Purpose for Submission:
New Device

C. Measurand:
Anti-nRNP/Sm, anti-Sm, anti-SS-A, anti-SS-B, anti-Scl-70, anti-Centromeres, anti-Jo-1, anti-Ribosomal P-proteins IgG antibodies (bundled)

D. Type of Test:
Qualitative, ELISA

E. Applicant:
EUROIMMUN US Inc.

F. Proprietary and Established Names:
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)

G. Regulatory Information:

1. Regulation section:
21 CFR§866.5110 – Antinuclear Antibody Immunological Test System

2. Classification:
Class II

3. Product code:

|  Device | Product Code  |
| --- | --- |
|  Anti-nRNP/Sm ELISA (IgG) | LKO, Anti-RNP Antibody, Antigen, Control  |
|  Anti-Sm ELISA (IgG) | LKP, Anti-Sm Antibody, Antigen and Control  |
|  Anti-SS-A ELISA (IgG) | LLL, Extractable antinuclear antibody, antigen and control  |
|  Anti-SS-B ELISA (IgG)  |   |
|  Anti-Scl-70 ELISA (IgG)  |   |
|  Anti-Jo-1 ELISA (IgG)  |   |
|  Anti-Centromeres ELISA (IgG) | LJM, Antinuclear antibody (Enzyme-labeled), antigen, control  |
|  Anti-Ribosomal P-Proteins ELISA (IgG) | MQA, Anti-ribosomal P antibodies  |

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4. Panel:

Immunology (82)

H. Intended Use:

1. Intended use(s):

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 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.

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.

2

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2. Indication(s) for use:
Same as Intended Use

3. Special conditions for use statement(s):
Prescription use only

4. Special instrument requirements:
Microwell plate reader capable of measuring OD at 450nm and at 620nm for dual wavelength readings.

I. Device Description:
The kit contains the following materials:

Device specific reagents: Microwell ELISA plate coated with antigen (nRNP-Sm, Sm, SS-A, SS-B, Scl-70, Jo-1, ribosomal P-proteins, centromeres, respectively)

Other reagents: 1 calibrator (20 RU/mL, IgG, human), positive and negative control (IgG, human), horseradish peroxidase (HRP)-labeled anti-human IgG conjugate, sample buffer concentrate, TMB chromogen/substrate solution and stop solution.

J. Substantial Equivalence Information:

1. Predicate device name(s) and 510(k) number(s):
INOVA Quanta Lite RNP ELISA (k922833)
INOVA Quanta Lite Sm ELISA (k922831)
INOVA Quanta Lite SS-A ELISA (k922830)
INOVA Quanta Lite SS-B ELISA (k922832)
INOVA Quanta Lite Scl-70 ELISA (k924898)
INOVA Quanta Lite Jo-1 ELISA (k926562)
INOVA Quanta Lite ribosomal P-proteins (k981237)
INOVA Quanta Lite Centromeres ELISA (k003959)

2. Comparison with predicate:

|  Similarities  |   |   |   |
| --- | --- | --- | --- |
|  Item |   | Device | Predicate  |
|  Intended Use | Anti-nRNP/Sm ELISA | Detection of IgG to nRNP/Sm | Same  |
|   |  Anti-Sm ELISA | Detection of IgG to Sm | Same  |
|   |  Anti-SS-A ELISA | Detection of IgG to SS-A | Same  |
|   |  Anti-SS-B ELISA | Detection of IgG to SS-B | Same  |
|   |  Anti-Scl-70 ELISA | Detection of IgG to Scl-70 | Same  |
|   |  Anti-Centromeres ELISA | Detection of IgG to centromeres | Same  |
|   |  Anti-Jo-1 ELISA | Detection of IgG to Jo-1 | Same  |
|   |  Anti-Ribosomal P-Proteins ELISA | Detection of IgG to ribosomal P-Protein | Same  |

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|  Similarities  |   |   |   |
| --- | --- | --- | --- |
|  Item |   | Device | Predicate  |
|  Antigen | Anti-RNP/Sm ELISA | Purified U1-nRNP/Sm | Same  |
|   |  Anti-Sm ELISA | Purified Sm | Same  |
|   |  Anti-SS-A ELISA | Purified SS-A | Same  |
|   |  Anti-SS-B ELISA | Purified SS-B | Same  |
|   |  Anti-Scl-70 ELISA | Purified Scl-70 | Same  |
|   |  Anti-Jo-1 ELISA | Purified Jo-1 | Same  |
|  Assay platform |   | 96-well microtiter plates | Same  |
|  Substrate |   | TMB | Same  |
|  Method |   | ELISA | Same  |
|  Differences  |   |   |   |
| --- | --- | --- | --- |
|  Item |   | Device | Predicate  |
|  Assay format |   | Qualitative | Semi-quantitative  |
|  Antigen | Anti-Centromeres ELISA | Recombinant centromeres protein B | Recombinant CENP-A and CENP-B  |
|   |  Anti-Ribosomal P-Proteins ELISA | Purified ribosomal P antigen | Synthetic ribosomal P antigen  |
|  Conjugate |   | Rabbit anti-human IgG labeled with HRP | Goat anti-human IgG labeled with HRP  |
|  Samples |   | Serum or plasma (EDTA, Li-heparin, Citrate) | Serum  |
|  Sample dilution |   | 1:201 | 1:101  |
|  Calibrator |   | 1 Calibrator: 20 RU/mL | None The low positive control is used for single point calibration  |
|  Control |   | 2 Controls: 1 positive and 1 negative | 3 Controls: 1 high positive, 1 low positive, 1 negative  |
|  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 Anti-Nuclear Antibody (ANA) Test System Premarket (510(k)) Submissions (January 22, 2009)

# L. Test Principle:

Patient samples are diluted 1:201 in sample buffer,  $100~\mu \mathrm{L}$  of each diluted patient sample and pre-diluted controls and calibrators are added to the antigen coated microtiter wells and

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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 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 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.

## M. Performance Characteristics (if/when applicable):

### 1. Analytical performance:

#### a. Precision/Reproducibility:

Intra- and inter-assay reproducibility: The reproducibility of each of eight tests was investigated for in the intra- and inter-assay using sera with values at different ratio. The intra-assay based on 20 determinations and the inter-assay based on 40 determinations performed in 5 days, 2 runs/day with 4 replicate/run. The precision data for all samples was analyzed to generate a summary of the qualitative reproducibility of the assay. The results are summarized below:

|  Anti-nRNP/Sm ELISA (IgG)  |   |   |   |   |
| --- | --- | --- | --- | --- |
|  Sample | Intra-assay (n=20) |   | Inter-assay (n=40)  |   |
|   |  Mean (Ratio) | % of positive | Mean (Ratio) | % of positive  |
|  1 | 5.1 | 100% | 5.2 | 100%  |
|  2 | 3.3 | 100% | 3.3 | 100%  |
|  3 | 1.9 | 100% | 1.8 | 100%  |
|  4 | 1.2 | 100% | 1.1 | 100%  |
|  5 | 0.8 | 0% | 0.8 | 0%  |
|  6 | 0.4 | 0% | 0.4 | 0%  |
|  Anti-Sm ELISA (IgG)  |   |   |   |   |
|  Sample | Intra-assay (n=20) |   | Inter-assay (n=40)  |   |
|   |  Mean (Ratio) | % of positive | Mean (Ratio) | % of positive  |
|  1 | 5.7 | 100% | 5.8 | 100%  |
|  2 | 3.7 | 100% | 3.9 | 100%  |
|  3 | 2.0 | 100% | 1.7 | 100%  |
|  4 | 1.2 | 100% | 1.1 | 100%  |
|  5 | 0.8 | 0% | 0.8 | 2.5%  |
|  6 | 0.4 | 0% | 0.4 | 0%  |
|  Anti-SS-A ELISA (IgG)  |   |   |   |   |
|  Sample | Intra-assay (n=20) |   | Inter-assay (n=40)  |   |
|   |  Mean (Ratio) | % of positive | Mean (Ratio) | % of positive  |
|  1 | 6.1 | 100% | 6.0 | 100%  |
|  2 | 4.2 | 100% | 4.0 | 100%  |
|  3 | 2.0 | 100% | 1.8 | 100%  |
|  4 | 1.3 | 100% | 1.2 | 100%  |
|  5 | 0.8 | 0% | 0.8 | 0%  |
|  6 | 0.3 | 0% | 0.3 | 0%  |

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|  Anti-SS-B ELISA (IgG)  |   |   |   |   |
| --- | --- | --- | --- | --- |
|  Sample | Intra-assay (n=20) |   | Inter-assay (n=40)  |   |
|   |  Mean (Ratio) | % of positive | Mean (Ratio) | % of positive  |
|  1 | 4.1 | 100% | 4.7 | 100%  |
|  2 | 2.9 | 100% | 3.2 | 100%  |
|  3 | 1.3 | 100% | 1.5 | 100%  |
|  4 | 0.8 | 0% | 1.1 | 100%  |
|  5 | 0.6 | 0% | 0.7 | 0%  |
|  6 | 0.4 | 0% | 0.5 | 0%  |
|  Anti-Scl-70 ELISA (IgG)  |   |   |   |   |
|  Sample | Intra-assay (n=20) |   | Inter-assay (n=40)  |   |
|   |  Mean (Ratio) | % of positive | Mean (Ratio) | % of positive  |
|  1 | 6.4 | 100% | 6.2 | 100%  |
|  2 | 4.2 | 100% | 4.2 | 100%  |
|  3 | 2.1 | 100% | 1.9 | 100%  |
|  4 | 1.4 | 100% | 1.2 | 100%  |
|  5 | 0.8 | 0% | 0.8 | 0%  |
|  6 | 0.4 | 0% | 0.3 | 0%  |
|  Anti-Centromeres ELISA (IgG)  |   |   |   |   |
|  Sample | Intra-assay (n=20) |   | Inter-assay (n=40)  |   |
|   |  Mean (Ratio) | % of positive | Mean (Ratio) | % of positive  |
|  1 | 4.5 | 100% | 4.7 | 100%  |
|  2 | 2.9 | 100% | 3.0 | 100%  |
|  3 | 1.5 | 100% | 1.5 | 100%  |
|  4 | 1.2 | 100% | 1.2 | 100%  |
|  5 | 0.6 | 0% | 0.7 | 0%  |
|  6 | 0.4 | 0% | 0.4 | 0%  |
|  Anti-Jo-1 ELISA (IgG)  |   |   |   |   |
|  Sample | Intra-assay (n=20) |   | Inter-assay (n=40)  |   |
|   |  Mean (Ratio) | % of positive | Mean (Ratio) | % of positive  |
|  1 | 6.4 | 100% | 6.0 | 100%  |
|  2 | 4.3 | 100% | 4.2 | 100%  |
|  3 | 1.8 | 100% | 2.2 | 100%  |
|  4 | 1.0 | 100% | 1.1 | 100%  |
|  5 | 0.8 | 0% | 0.8 | 0%  |
|  6 | 0.4 | 0% | 0.4 | 0%  |
|  Anti-ribosomal P-protein ELISA (IgG)  |   |   |   |   |
|  Sample | Intra-assay (n=20) |   | Inter-assay (n=40)  |   |
|   |  Mean (Ratio) | % of positive | Mean (Ratio) | % of positive  |
|  1 | 5.5 | 100% | 5.7 | 100%  |
|  2 | 3.6 | 100% | 3.8 | 100%  |
|  3 | 1.9 | 100% | 1.9 | 100%  |
|  4 | 1.0 | 100% | 1.1 | 100%  |
|  5 | 0.6 | 0% | 0.7 | 0%  |
|  6 | 0.4 | 0% | 0.4 | 0%  |

Lot-to-Lot reproducibility: The lot to lot reproducibility was also investigated by using various lots with multiple samples at different ratio. The results are

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summarized in the following tables.

|  Anti-nRNP/Sm ELISA (IgG)  |   |   |   |   |   |
| --- | --- | --- | --- | --- | --- |
|  Sample No. | N=6 (3 lots x 2 runs) |   | Sample No. | N=11 (11 lots x 1 run)  |   |
|   |  Mean ratio | % of Positive |   | Mean ratio | % of Positive  |
|  1 | 3.6 | 100% | 7 | 0.2 | 0%  |
|  2 | 2.5 | 100% | 8 | 3.4 | 100%  |
|  3 | 5.4 | 100% | 9 | 5.2 | 100%  |
|  4 | 0.4 | 0% | 10 | 6.6 | 100%  |
|  5 | 0.9 | 0% | 11 | 8.5 | 100%  |
|  6 | 1.1 | 100% |  |  |   |
|  Anti-Sm ELISA (IgG)  |   |   |   |   |   |
|  Sample No. | N=6 (3 lots x 2 runs) |   | Sample No. | N=n (n lots x 1 run)  |   |
|   |  Mean ratio | % of Positive |   | Mean ratio (N) | % of Positive  |
|  1 | 3.6 | 100% | 7 | 0.1 (9) | 0%  |
|  2 | 3.4 | 100% | 8 | 1.9 (10) | 100%  |
|  3 | 3.1 | 100% | 9 | 3.4 (10) | 100%  |
|  4 | 0.3 | 0% | 10 | 5.7 (9) | 100%  |
|  5 | 0.9 | 0% | 11 | 8.3 (10) | 100%  |
|  6 | 1.2 | 100% |  |  |   |
|  Anti-SS-A ELISA (IgG)  |   |   |   |   |   |
|  Sample No. | N=6 (3 lots x 2 runs) |   | Sample No. | N=n lots x 1 run  |   |
|   |  Mean Ratio | % of Positive |   | Mean Ratio (N) | % of Positive  |
|  1 | 4.6 | 100% | 7 | 0.1 (10) | 0%  |
|  2 | 5.6 | 100% | 8 | 1.6 (11) | 100%  |
|  3 | 5.6 | 100% | 9 | 2.7 (10) | 100%  |
|  4 | 0.3 | 0% | 10 | 4.6 (9) | 100%  |
|  5 | 0.9 | 0% | 11 | 7.8 (11) | 100%  |
|  6 | 1.1 | 100% |  |  |   |
|  Anti-SS-B ELISA (IgG)  |   |   |   |   |   |
|  Sample No. | N=6 (3 lots x 2 runs) |   | Sample No. | N=11 (11 lots x 1 run)  |   |
|   |  Mean Ratio | % of Positive |   | Mean Ratio | % of Positive  |
|  1 | 3.9 | 100% | 7 | 2.6 | 100%  |
|  2 | 3.9 | 100% | 8 | 4.2 | 100%  |
|  3 | 3.8 | 100% | 9 | 7.7 | 100%  |
|  4 | 0.3 | 0% |  |  |   |
|  5 | 0.9 | 0% |  |  |   |
|  6 | 1.2 | 100% |  |  |   |

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|  Anti-Scl-70 ELISA (IgG)  |   |   |   |   |   |
| --- | --- | --- | --- | --- | --- |
|  Sample No. | N=6 (3 lots x 2 runs) |   | Sample No. | N=n (n lots x 1 run)  |   |
|   |  Mean Ratio | % of Positive |   | Mean Ratio (N) | % of Positive  |
|  1 | 4.5 | 100% | 7 | 3.0 (11) | 100%  |
|  2 | 4.7 | 100% | 8 | 4.8 (10) | 100%  |
|  3 | 3.6 | 100% | 9 | 6.2 (11) | 100%  |
|  4 | 0.3 | 0% |  |  |   |
|  5 | 0.9 | 0% |  |  |   |
|  6 | 1.1 | 100% |  |  |   |
|  Anti-Centromeres ELISA (IgG)  |   |   |   |   |   |
|  Sample No. | N=6 (3 lots x 2 runs) |   | Sample No. | N=n (n lots x 1 run)  |   |
|   |  Mean Ratio | % of Positive |   | Mean Ratio (N) | % of Positive  |
|  1 | 4.1 | 100% | 7 | 2.8 (8) | 100%  |
|  2 | 4.9 | 100% | 8 | 6.2 (10) | 100%  |
|  3 | 3.9 | 100% | 9 | 6.8 (11) | 100%  |
|  4 | 0.3 | 0% | 10 | 8.4 (10) | 100%  |
|  5 | 0.9 | 0% |  |  |   |
|  6 | 1.1 | 100% |  |  |   |
|  Anti-Jo-1 ELISA (IgG)  |   |   |   |   |   |
|  Sample No. | N=6 (3 lots x 2 runs) |   | Sample No. | N=n (n lots x 1 run)  |   |
|   |  Mean Ratio | % of Positive |   | Mean Ratio (N) | % of Positive  |
|  1 | 5.1 | 100% | 7 | 2.6 (10) | 100%  |
|  2 | 3.9 | 100% | 8 | 4.0 (11) | 100%  |
|  3 | 2.0 | 100% | 9 | 7.4 (11) | 100%  |
|  4 | 0.3 | 0% | 10 | 8.2 (11) | 100%  |
|  5 | 0.9 | 0% |  |  |   |
|  6 | 1.1 | 100% |  |  |   |
|  Anti-Ribosomal P-protein ELISA (IgG)  |   |   |   |   |   |
|  Sample No. | N=6 (3 lots x 2 runs) |   | Sample No. | N=11 (11 lots x 1 run)  |   |
|   |  Mean Ratio | % of Positive |   | Mean Ratio | % of Positive  |
|  1 | 3.2 | 100% | 7 | 4.8 | 100%  |
|  2 | 4.5 | 100% | 8 | 5.2 | 100%  |
|  3 | 2.2 | 100% | 9 | 7.6 | 100%  |
|  4 | 0.3 | 0% |  |  |   |
|  5 | 0.9 | 0% |  |  |   |
|  6 | 1.1 | 100% |  |  |   |

# b. Linearity/assay reportable range:

Not applicable

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

Traceability: There is no reference standard for these analytes. The reactivity of each of eight devices was verified using the ANA reference panel of the CDC (Centers for

{8}

Disease Control and Prevention, Atlanta, USA). The ANA reference panel include native DNA, SS-B, RNP/SS-B/SS-A, RNP, Sm, U3-RNP, SS-A, Centromeres, Scl-70, Jo-1, PM-Scl, and ribosomal P-proteins. All eight devices showed the expected reactivity against the reference panel.

**Calibrator and Controls:** The cut-off calibrator and controls are derived from human serum and match pre-specified performance criteria. Each lot of cut-off calibrator and controls are traceable to a master lot.

**Stability:** Real time stability studies were conducted using three production lots of kit reagents. The results support a shelf life claim of 12 months for all components when stored at  $2 - 8^{\circ}\mathrm{C}$ . The opened reagents are stable for 6 months when stored at  $2 - 8^{\circ}\mathrm{C}$ . The reconstituted Wash Buffer is stable for up to 28 days.

# d. Detection limit:

Not applicable

# e. Analytical specificity:

Cross reactivity: To investigate the quality of the antigen coated on the plates to ensure a high specificity of the ELISA, cross reactivity was investigative using a panel of 30 sera containing antibodies serologically positive against the known antigens. The panel of antibodies used to test antigen coated for each device listed in the following table:

|  Device | Panel of antibodies used for Cross Reactivity Study  |
| --- | --- |
|  Anti-nRNP/Sm | Anti-Ribosomal P-protein, SS-A, SS-B, Scl-70, Jo-1, centromeres, Rubella virus, Measles virus, Herpes simplex virus type 1 and Borrelia burgdorferi  |
|  Anti-Sm | Anti-Ribosomal P-protein, nRNP/Sm, SS-A, SS-B, Scl-70, Jo-1, centromeres, Rubella virus, Measles virus, Herpes simplex virus type 1 and Borrelia burgdorferi  |
|  Anti-SS-A | Anti-Ribosomal P-protein, nRNP/Sm, Sm, Scl-70, Jo-1, centromeres, Rubella virus, Measles virus, Herpes simplex virus type 1 and Borrelia burgdorferi  |
|  Anti-SS-B | Anti-Ribosomal P-protein, nRNP/Sm, Sm, SS-A, Scl-70, Jo-1, centromeres, Rubella virus, Measles virus, Herpes simplex virus type 1 and Borrelia burgdorferi  |
|  Anti-Scl-70 | Anti-Ribosomal P-protein, nRNP/Sm, Sm, SS-A, SS-B, Jo-1, centromeres, Rubella virus, Measles virus, Herpes simplex virus type 1 and Borrelia burgdorferi  |
|  Anti-Centromeres | Anti-Ribosomal P-protein, nRNP/Sm, Sm, SS-A, SS-B, Scl-70, Jo-1, Rubella virus, Measles virus, Herpes simplex virus type 1 and Borrelia burgdorferi  |

{9}

10

|  Device | Panel of antibodies used for Cross Reactivity Study  |
| --- | --- |
|  Anti-Jo-1 | Anti-Ribosomal P-protein, nRNP/Sm, Sm, SS-A, SS-B, Scl-70, centromeres, Rubella virus, Measles virus, Herpes simplex virus type 1 and Borrelia burgdorferi  |
|  Anti-ribosomal P-proteins | Anti-nRNP/Sm, Sm, SS-A, SS-B, Scl-70, Jo-1, centromeres, Rubella virus, Measles virus, Herpes simplex virus type 1 and Borrelia burgdorferi  |

No cross reactivity is observed with the tested antibody panel for each device.

**Endogenous interference**: For each device, interference testing was performed by using 5 serum samples with various ratio of each analyte (negative, positive and near to cut-off). Samples were spiked with potential interfering substances, hemoglobin (250, 500, and 1000 mg/dL), triglycerides (500, 1000, 2000 mg/dL), bilirubin (10, 20, and 40 mg/dL) and rheumatoid factor (RF) (500 IU/mL). Recoveries of all spiked sample/interferent combinations compared to the measurements for unspiked samples were calculated. 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 RF,

**f. Assay cut-off**:

For each device, the assay cut-off is OD ratio of 1.0. Results ≥ 1.0 are positive, and results < 1.0 are negative.

**2. Comparison studies**:

**a. Method comparison with predicate device**:

**Anti-nRNP/Sm ELISA (IgG)**: A total of 287 samples were tested with EUROIMMUN Anti-nRNP/Sm ELISA (IgG) and predicate device. The samples were from 52 mixed connective tissue disease (MCTD), 69 systemic lupus erythematosus (SLE), 51 Sjögren’s syndrome, 15 systemic sclerosis, 15 fibromyalgia, 35 rheumatoid arthritis (RA), 30 borreliosis, and 20 healthy individuals. Anti-nRNP/Sm antibodies are expected in either MCTD or SLE. The other disease groups serve as control cohorts. The results are shown in the table below. Of the 8 discrepant samples, one was from a MCTD patient and the other 7 were from controls.

|   | Inova Quanta Lite RNP ELISA  |   |   |   |
| --- | --- | --- | --- | --- |
|   |   |  Positive | Negative | Total  |
|  EUROIMMUN
Anti-nRNP/Sm
ELISA (IgG) | Positive | 83 | 0 | 83  |
|   |  Negative | 8 | 196 | 204  |
|   |  Total | 91 | 196 | 287  |

Positive agreement: 91.2% (83/91) (95% CI: 83.4 – 96.1%)

Negative agreement: 100.0% (196/196) (95% CI: 98.1 – 100%)

Overall agreement: 97.2% (279/287) (95% CI: 94.6 – 98.8%)

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Anti-Sm ELISA (IgG): A total of 294 clinical samples were tested with EUROIMMUN Anti-nRNP/Sm ELISA (IgG) and predicate device. The samples were from 128 SLE, 51 Sjögren's syndrome, 15 systemic sclerosis, 15 fibromyalgia, 35 RA, 30 borreliosis and 20 healthy individuals. The results are shown in the table below. Anti-Sm antibodies are expected in SLE. The other disease groups serve as control cohorts. 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.

|   | Inova Quanta Lite Sm ELISA  |   |   |   |
| --- | --- | --- | --- | --- |
|   |   |  Positive | Negative | Total  |
|  EUROIMMUN Anti-Sm ELISA (IgG) | Positive | 37 | 5 | 42  |
|   |  Negative | 7 | 245 | 252  |
|   |  Total | 44 | 250 | 294  |

Positive agreement:  84.1% (37/44)  (95% CI: 69.9 - 93.4%)

Negative agreement:  98.0% (245/250)  (95% CI: 95.4 - 99.3%)

Overall agreement:  97.2% (279/287)  (95% CI: 94.6 - 98.8%)

## Anti-SS-A ELISA IgG:

A total of 305 samples were tested with EUROIMMUN Anti-SS-A ELISA (IgG) and predicate device. The samples were from 63 SLE, 77 Sjögren's syndrome, 23 systemic sclerosis, 1 systemic sclerosis/ Sjögren's syndrome, 15 fibromyalgia, 26 myositis, 35 RA, 30 borreliosis, 20 healthy individuals. Anti-SS-A antibodies are expected in either Sjögren's syndrome or SLE. The other disease groups serve as control cohorts. Of the 9 discrepant samples negative in the EUROIMMUN test, 2 were from patients with Sjögren'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.

|   | Inova Quanta Lite SS-A ELISA  |   |   |   |
| --- | --- | --- | --- | --- |
|   |   |  Positive | Negative | Total  |
|  EUROIMMUN Anti-SS-A ELISA (IgG) | Positive | 116 | 3 | 119  |
|   |  Negative | 9 | 177 | 186  |
|   |  Total | 125 | 180 | 305  |

Positive agreement:  92.8% (116/125)  (95% CI: 86.8 - 96.7%)

Negative agreement:  98.3% (177/180)  (95% CI: 95.2 - 99.7%)

Overall agreement:  96.1% (293/305)  (95% CI: 93.2 - 98.0%)

## Anti-SS-B IgG:

A total of 275 samples were tested with EUROIMMUN Anti-SS-B ELISA (IgG) and predicate device. The samples were from 57 SLE, 64 Sjögren's syndrome, 23 systemic sclerosis, 1 systemic sclerosis/Sjögren's syndrome, 4 SLE/ Sjögren's syndrome, 15 fibromyalgia, 26 myositis, 35 RA, 30 borreliosis and 20 healthy individuals. Anti-SS-B antibodies are expected in either Sjögren's syndrome or SLE. The other disease groups serve as control cohorts. Of 9 discrepant samples negative in the EUROIMMUN test, 2 were from Sjögren's syndrome patients and 1 were from

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a SLE patient, the other 6 were from controls. All 3 discrepant samples positive in the ERUOIMMUN test were from SLE patients.

|   | Inova Quanta Lite SS-A ELISA  |   |   |   |
| --- | --- | --- | --- | --- |
|   |   |  Positive | Negative | Total  |
|  EUROIMMUN Anti-SS-A ELISA (IgG) | Positive | 116 | 3 | 119  |
|   |  Negative | 9 | 177 | 186  |
|   |  Total | 125 | 180 | 305  |

Positive agreement:  $84.1\%$  (37/44)  $(95\% \mathrm{CI}:69.9 - 93.4\%)$

Negative agreement:  $98.0\%$  (245/250)  $(95\% \mathrm{CI}: 95.4 - 99.3\%)$

Overall agreement:  $97.2\%$  (279/287)  $(95\% \mathrm{CI}: 94.6 - 98.8\%)$

# Anti-Scl-70 IgG:

A total of 309 samples were tested with EUROIMMUN Anti-Scl-70 ELISA (IgG) and predicate device. The samples were from 158 systemic sclerosis, 51 Sjögren's syndrome, 15 fibromyalgia, 35 RA, 30 borreliosis and 20 healthy individuals. Anti-Scl-70 antibodies are expected in systemic sclerosis. The other disease groups serve as control cohorts.

|   | Inova Quanta Lite Scl-70 ELISA  |   |   |   |
| --- | --- | --- | --- | --- |
|   |   |  Positive | Negative | Total  |
|  EUROIMMUN Anti-Scl-70 ELISA (IgG) | Positive | 125 | 0 | 125  |
|   |  Negative | 0 | 184 | 184  |
|   |  Total | 125 | 184 | 309  |

Positive agreement:  $100.0\%$  (125/125)  $(95\% \mathrm{CI}: 97.1 - 100\%)$

Negative agreement:  $100.0\%$  (184/184)  $(95\% \mathrm{CI}: 98.0 - 100\%)$

Overall agreement:  $100.0\%$  (309/309)  $(95\% \mathrm{CI}: 98.9 - 100\%)$

# Anti-Centromeres ELISA IgG:

A total of 297 samples were tested with EUROIMMUN Anti-Centromeres ELISA (IgG) and predicate device. The samples were from 144 systemic sclerosis, 2 CREST, 51 Sjögren's syndrome, 15 fibromyalgia, 35 RA, 30 borreliosis and 20 healthy individuals. Anti-Centromeres antibodies are expected in systemic sclerosis. The other disease groups serve as control cohorts. All of the 5 discrepant samples were from control groups.

|   | Inova Quanta Lite Centromere ELISA  |   |   |   |
| --- | --- | --- | --- | --- |
|   |   |  Positive | Negative | Total  |
|  EUROIMMUN Anti-Centromeres ELISA (IgG) | Positive | 73 | 0 | 73  |
|   |  Negative | 5 | 219 | 224  |
|   |  Total | 78 | 219 | 297  |

Positive agreement:  $93.6\%$  (73/78)  $(95\% \mathrm{CI}: 85.7 - 97.9\%)$

Negative agreement:  $100.0\%$  (219/219)  $(95\% \mathrm{CI}: 98.3 - 100\%)$

Overall agreement:  $98.3\%$  (292/297)  $(95\% \mathrm{CI}: 96.1 - 99.5\%)$

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13

# Anti-Jo-1 ELISA IgG:

A total of 297 samples were tested with EUROIMMUN Anti-Jo-1 ELISA (IgG) and predicate device. The samples were from 143 myositis, 3 Polymyositis/systemic sclerosis, 51 Sjögren’s syndrome, 15 fibromyalgia, 35 RA, 30 borreliosis and 20 healthy individuals. Anti-Jo-1 antibodies are expected in myositis. The other disease groups serve as control cohorts. Both discrepant samples were from myositis patients.

|   | Inova Quanta Lite Jo-1 ELISA  |   |   |   |
| --- | --- | --- | --- | --- |
|   |   |  Positive | Negative | Total  |
|  EUROIMMUN
Anti-Jo-1
ELISA (IgG) | Positive | 64 | 1 | 65  |
|   |  Negative | 1 | 231 | 232  |
|   |  Total | 65 | 232 | 297  |

Positive agreement: 98.5% (64/65) (95% CI: 91.7 – 100%)
Negative agreement: 99.6% (231/232) (95% CI: 97.6 – 100%)
Overall agreement: 99.3% (295/297) (95% CI: 97.6 – 99.9%)

# Anti-ribosomal P-proteins ELISA (IgG):

A total of 243 samples were tested with EUROIMMUN Anti-ribosomal P-proteins ELISA (IgG) and predicate device. The samples were from 90 SLE, 1 SLE/Sjögren’s syndrome, 1 cutaneous lupus erythematosus (CLE), 51 Sjögren’s syndrome, 15 fibromyalgia, 35 RA, 30 borreliosis and 20 healthy individuals. Anti-ribosomal P-proteins antibodies are expected in SLE. The other disease groups serve as control cohorts. All of the 10 discrepant samples were from patients with SLE, SLE/Sjögren’s syndrome, and CLE.

|   | Inova Quanta Lite Ribosomal P ELISA  |   |   |   |
| --- | --- | --- | --- | --- |
|   |   |  Positive | Negative | Total  |
|  EUROIMMUN
Anti-ribosomal
P-proteins
ELISA (IgG) | Positive | 28 | 10 | 38  |
|   |  Negative | 0 | 205 | 205  |
|   |  Total | 28 | 215 | 243  |

Positive agreement: 100.0% (28/28) (95% CI: 87.7 – 100%)
Negative agreement: 95.3% (205/215) (95% CI: 91.6 – 97.7%)
Overall agreement: 95.9% (233/243) (95% CI: 92.6 – 98.0%)

## b. Matrix comparison:

The matrix comparison was evaluated using sample pairs of serum and corresponding plasma (EDTA, Li-heparin, and Citrate) tested with each device. The results are summarized in the following table:

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|   | Matrix compared to serum  |   |   |
| --- | --- | --- | --- |
|   |  EDTA | Li-Heparin | Citrate  |
|  Anti-nRNP/Sm ELISA  |   |   |   |
|  N | 15 | 15 | 15  |
|  Ratio range (serum) | 0.3 – 10.4 | 0.3 – 10.4 | 0.3 – 10.4  |
|  Ratio range (plasma) | 0.3 – 10.0 | 0.3 – 11.1 | 0.3 – 11.5  |
|  % recovery | 91 – 117% | 92 – 114% | 93 – 120%  |
|  Anti-Sm ELISA  |   |   |   |
|  N | 16 | 16 | 16  |
|  Ratio range (serum) | 0.2 – 9.9 | 0.2 – 9.9 | 0.2 – 9.9  |
|  Ratio range (plasma) | 0.3 – 9.6 | 0.2 – 9.4 | 0.2 – 9.6  |
|  % recovery: | 94 – 115% | 90 – 111% | 89 – 118%  |
|  Anti-SS-A ELISA  |   |   |   |
|  N | 22 | 22 | 22  |
|  Ratio range (serum) | 0.2 – 9.8 | 0.2 – 9.8 | 0.2 – 9.8  |
|  Ratio range (plasma) | 0.2 – 9.9 | 0.2 – 9.9 | 0.2 – 9.8  |
|  % recovery: | 90 – 106% | 85 – 117% | 89 – 116%  |
|  Anti-SS-B ELISA  |   |   |   |
|  N | 16 | 15 | 15  |
|  Ratio range (serum) | 0.3 – 9.4 | 0.3 – 9.4 | 0.3 – 9.4  |
|  Ratio range (plasma) | 0.2 – 10.0 | 0.3 – 10.0 | 0.2 – 9.4  |
|  % recovery: | 78 – 116% | 98 – 116% | 81 – 110%  |
|  Anti-Scl-70 ELISA  |   |   |   |
|  N | 18 | 18 | 18  |
|  Ratio range (serum) | 0.3 – 9.7 | 0.3 – 9.7 | 0.3 – 9.7  |
|  Ratio range (plasma) | 0.3 – 9.7 | 0.3 – 9.8 | 0.4 – 9.7  |
|  % recovery: | 86 – 107% | 92 – 113% | 87 – 106%  |
|  Anti-Centromeres ELISA  |   |   |   |
|  N | 16 | 16 | 16  |
|  Ratio range (serum) | 0.3 – 9.4 | 0.3 – 9.4 | 0.3 – 9.4  |
|  Ratio range (plasma) | 0.3 – 9.2 | 0.3 – 8.4 | 0.3 – 9.2  |
|  % recovery: | 84 – 108% | 83 – 101% | 88 – 107%  |
|  Anti-Jo-1 ELISA  |   |   |   |
|  N | 15 | 15 | 15  |
|  Ratio range (serum) | 0.3 – 9.2 | 0.3 – 9.2 | 0.3 – 9.2  |
|  Ratio range (plasma) | 0.3 – 8.7 | 0.4 – 8.5 | 0.3 – 8.9  |
|  % recovery: | 85 – 117% | 87 – 113% | 94 – 114%  |
|  Anti-Ribosomal P-protein ELISA  |   |   |   |
|  N | 17 | 17 | 17  |
|  Ratio range (serum) | 0.3 – 9.6 | 0.3 – 9.6 | 0.3 – 9.6  |
|  Ratio range (plasma) | 0.3 – 9.4 | 0.3 – 9.7 | 0.3 – 9.5  |
|  % recovery: | 79 – 107% | 80 – 107% | 85 – 107%  |

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# 3. Clinical studies:

# a. Clinical Sensitivity/Clinical Specificity:

Anti-nRNP/Sm IgG: A total of 1046 clinically characterized samples (65 MCTD, 404 SLE, and 577 from other control diseases) were evaluated for clinical sensitivity and specificity with EUROIMMUN anti-nRNP/Sm ELISA (IgG). The results are summarized in the following table.

Anti-nRNP/Sm ELISA (IgG):

|  Clinial Sensitivity  |   |   |   |   |
| --- | --- | --- | --- | --- |
|  Panel | N | Positive | % Positive | 95% CI  |
|  MCTD | 65 | 65 | 100.0% | 94.5 – 100.0%  |
|  SLE | 404 | 94 | 23.3% | 19.2 – 27.7%  |
|  Clinical Specificity  |   |   |   |   |
|  Panel | N | Negative | % Negative | 95% CI  |
|  RA | 164 | 164 | 100.0% | 97.8 – 100.0%  |
|  Polymyositis/Dermatomyositis | 151 | 143 | 94.7% | 89.8 – 97.7%  |
|  Systemic sclerosis | 81 | 81 | 100.0% | 95.5 – 100.0%  |
|  Sjögren's syndrome | 88 | 86 | 97.7% | 92.0 – 99.7%  |
|  Other autoimmune diseases* | 63 | 62 | 98.4% | 91.5 – 100.0%  |
|  Borreliosis | 30 | 30 | 100.0% | 88.4 – 100.0%  |
|  Total | 577 | 566 | 98.1% | 96.6 – 99.0%  |

* Samples include: autoimmune hepatitis (n=8), primary biliary cirrhosis (n=9), Grave's disease (n=12), Hashimoto (n=11), celiac disease (n=11), Diabetes Type I (n=12)

Anti-Sm ELISA IgG: A total of 1038 clinically characterized samples (414 SLE, and 626 other control diseases) were evaluated for clinical sensitivity and specificity with EUROIMMUN anti-Sm ELISA (IgG). The results are summarized in the following table:

Anti-Sm ELISA (IgG):

|  Clinial Sensitivity  |   |   |   |   |
| --- | --- | --- | --- | --- |
|  Panel | N | Positive | % Positive | 95% CI  |
|  SLE | 414 | 47 | 11.4% | 8.5 – 14.8%  |
|  Clinical Specificity  |   |   |   |   |
|  Panel | N | Negative | % Negative | 95% CI  |
|  RA | 164 | 164 | 100.0% | 97.8 – 100.0%  |
|  Systemic sclerosis | 81 | 81 | 100.0% | 95.5 – 100.0%  |
|  Sjögren's syndrome | 88 | 88 | 100.0% | 95.9 – 100.0%  |
|  Polymyositis/Dermatomyositis | 151 | 151 | 100.0% | 97.6 – 100.0%  |

{15}

# Anti-SS-A ELISA: A total of 1026 clinically characterized samples (88 from Sjögren's syndrome patients, 404 from SLE, and 534 from control diseases) were evaluated for clinical sensitivity and specificity with EUROIMMUN anti-SS-A ELISA (IgG). The results are summarized in the following table:

Anti-SS-A ELISA (IgG):

|  Clinical Sensitivity  |   |   |   |   |
| --- | --- | --- | --- | --- |
|  Panel | N | Positive | % Positive | 95% CI  |
|  Sjögren's syndrome | 88 | 65 | 73.9% | 63.4 – 82.7%  |
|  SLE | 404 | 164 | 40.6% | 35.8 – 45.6%  |
|  Clinical Specificity  |   |   |   |   |
|  Panel | N | Negative | % Negative | 95% CI  |
|  Systemic sclerosis | 81 | 75 | 92.6% | 84.6 – 97.2%  |
|  Polymyositis/Dermatomyositis | 151 | 138 | 91.4% | 85.7 – 95.3%  |
|  RA | 164 | 159 | 97.0% | 93.0 – 99.0%  |
|  MCTD | 45 | 41 | 91.1% | 78.8 – 97.5%  |
|  Other autoimmune diseases* | 63 | 63 | 100.0% | 94.3 – 100.0%  |
|  Borreliosis | 30 | 30 | 100.0% | 88.4 – 100.0%  |
|  Total | 534 | 506 | 94.8% | 92.5 – 96.5%  |

* Samples include: autoimmune hepatitis (n=8), primary biliary cirrhosis (n=9), Grave's disease (n=12), Hashimoto (n=11), celiac disease (n=11), Diabetes Type I (n=12)

Anti-SS-B IgG: A total of 1026 clinically characterized samples (88 from Sjögren's syndrome patients, 404 from SLE, and 534 from control diseases) were evaluated for clinical sensitivity and specificity with EUROIMMUN anti-SS-B ELISA (IgG). The results are summarized in the following table:

Anti-SS-B ELISA (IgG):

|  Clinical Sensitivity  |   |   |   |   |
| --- | --- | --- | --- | --- |
|  Panel | N | Positive | % Positive | 95% CI  |
|  Sjögren's syndrome | 88 | 35 | 39.8% | 29.5 – 50.8%  |
|  SLE | 404 | 56 | 13.9% | 10.6 – 17.6%  |
|  Clinical Specificity  |   |   |   |   |
|  Panel | N | Negative | % Negative | 95% CI  |

{16}

|  RA | 164 | 163 | 99.4% | 96.6 – 100.0%  |
| --- | --- | --- | --- | --- |
|  Systemic sclerosis | 81 | 77 | 95.1% | 87.8 – 98.6%  |
|  Polymyositis/Dermatomyositis | 151 | 149 | 98.7% | 95.3 – 99.8%  |
|  MCTD | 45 | 42 | 93.3% | 81.7 – 98.6%  |
|  Other autoimmune diseases* | 63 | 63 | 100.0% | 94.3 – 100.0%  |
|  Borreliosis | 30 | 30 | 100.0% | 88.4 – 100.0%  |
|  Total | 534 | 524 | 98.1% | 96.6 – 99.1%  |

* Samples include: autoimmune hepatitis (n=8), primary biliary cirrhosis (n=9), Grave's disease (n=12), Hashimoto (n=11), celiac disease (n=11), Diabetes Type I (n=12)

Anti-Scl-70 ELISA: A total of 909 clinically characterized samples (280 from systemic sclerosis patients and 629 from control groups) were evaluated for clinical sensitivity and specificity with EUROIMMUN anti-Scl-70 ELISA (IgG). The results are summarized in the following table:

Anti-Scl-70 ELISA (IgG):

|  Clinical Sensitivity  |   |   |   |   |
| --- | --- | --- | --- | --- |
|  Panel | N | Positive | % Positive | 95% CI  |
|  Systemic sclerosis | 280 | 65 | 23.2% | 18.4 – 28.6%  |
|  - Diffuse systemic sclerosis | 96 | 57 | 59.4% | 48.9 – 69.3%  |
|  - Limited systemic sclerosis | 113 | 6 | 5.3% | 2.0 – 11.2%  |
|  Clinical Specificity  |   |   |   |   |
|  Panel | N | Negative | % Negative | 95% CI  |
|  SLE | 213 | 213 | 100.0% | 98.3 – 100.0%  |
|  Polymyositis/Dermatomyositis | 26 | 26 | 100.0% | 86.8 – 100.0%  |
|  RA | 164 | 163 | 99.4% | 96.6 – 100.0%  |
|  Sjögren's syndrome | 88 | 88 | 100.0% | 95.9 – 100.0%  |
|  MCTD | 45 | 45 | 100.0% | 92.1 – 100.0%  |
|  Other autoimmune diseases* | 63 | 63 | 100.0% | 94.3 – 100.0%  |
|  Borreliosis | 30 | 30 | 100.0% | 88.4 – 100.0%  |
|  Total | 629 | 628 | 99.8% | 99.1 – 100.0%  |

* Samples include: autoimmune hepatitis (n=8), primary biliary cirrhosis (n=9), Grave's disease (n=12), Hashimoto (n=11), celiac disease (n=11), Diabetes Type I (n=12)

Anti-Centromeres ELISA: A total of 877 clinically characterized samples (280 from systemic sclerosis patients and 597 from control groups) were investigated for clinical sensitivity and specificity with EUROIMMUN Anti-Centromeres ELISA (IgG). The results are summarized in the following table:

Anti-Centromeres ELISA (IgG):

{17}

|  Clinical Sensitivity  |   |   |   |   |
| --- | --- | --- | --- | --- |
|  Panel | N | Positive | % Positive | 95% CI  |
|  Systemic sclerosis | 280 | 105 | 37.5% | 31.8 – 43.5%  |
|  - Diffuse systemic sclerosis | 96 | 7 | 7.3% | 3.0 – 14.4%  |
|  - Limited systemic sclerosis | 113 | 84 | 74.3% | 65.3 – 82.1%  |
|  Clinical Specificity  |   |   |   |   |
|  Panel | N | Negative | % Negative | 95% CI  |
|  SLE | 181 | 180 | 99.4% | 97.0 – 100.0%  |
|  Polymyositis/Dermatomyositis | 26 | 26 | 100.0% | 86.8 – 100.0%  |
|  RA | 164 | 163 | 99.4% | 96.6 – 100.0%  |
|  Sjögren’s syndrome | 88 | 85 | 96.6% | 90.4 – 99.3%  |
|  MCTD | 45 | 44 | 97.8% | 88.2 – 99.9%  |
|  Other autoimmune diseases* | 63 | 63 | 100.0% | 94.3 – 100.0%  |
|  Borreliosis | 30 | 30 | 100.0% | 88.4 – 100.0%  |
|  Total | 597 | 591 | 99.0% | 97.8 – 99.6%  |

* Samples include: autoimmune hepatitis (n=8), primary biliary cirrhosis (n=9), Grave’s disease (n=12), Hashimoto (n=11), celiac disease (n=11), Diabetes Type I (n=12)

Anti-Jo-1 IgG: A total of 876 clinically characterized samples (177 from myositis patients and 699 from control groups) were evaluated for clinical sensitivity and specificity with EUROIMMUN Anti-Jo-1 ELISA (IgG). The results are summarized in the following table:

Anti-Jo-1 ELISA (IgG):

|  Clinical Sensitivity  |   |   |   |   |
| --- | --- | --- | --- | --- |
|  Panel | N | Positive | % Positive | 95% CI  |
|  Polymyositis/ Dermatomyositis | 177 | 33 | 18.6% | 13.2 – 25.2%  |
|  Clinical Specificity  |   |   |   |   |
|  Panel | N | Negative | % Negative | 95% CI  |
|  SLE | 213 | 211 | 99.1% | 96.6 – 99.9%  |
|  RA | 164 | 163 | 99.4% | 96.6 – 100.0%  |
|  Systemic sclerosis | 81 | 81 | 100.0% | 95.5 – 100.0%  |
|  Sjögren’s syndrome | 88 | 88 | 100.0% | 95.9 – 100.0%  |
|  MCTD | 45 | 45 | 100.0% | 92.1 – 100.0%  |
|  Fibromyalgia | 15 | 15 | 100.0% | 78.2 – 100.0%  |
|  Other autoimmune diseases | 63 | 63 | 100.0% | 94.3 – 100.0%  |
|  Borreliosis | 30 | 30 | 100.0% | 88.4 – 100.0%  |
|  Total | 699 | 696 | 99.6% | 98.8 – 99.9%  |

* Samples include: autoimmune hepatitis (n=8), primary biliary cirrhosis (n=9), Grave’s disease (n=12), Hashimoto (n=11), celiac disease (n=11), Diabetes Type I (n=12)

{18}

Anti-ribosomal P-proteins IgG: A total of 876 clinically characterized samples (376 from SLE patients and 500 from control groups) were evaluated for clinical sensitivity and specificity with EUROIMMUN Anti-ribosomal P-proteins ELISA (IgG). The results are summarized in the following table:

Anti-ribosomal P-proteins ELISA (IgG):

|  Clinical Sensitivity  |   |   |   |   |
| --- | --- | --- | --- | --- |
|  Panel | N | Positive | % Positive | 95% CI  |
|  SLE | 376 | 20 | 5.3% | 3.3 – 8.1%  |
|  Clinical Specificity  |   |   |   |   |
|  Panel | N | Negative | % Negative | 95% CI  |
|  Polymyositis/Dermatomyositis | 151 | 149 | 98.7% | 95.3 – 99.8%  |
|  RA | 90 | 90 | 100.0% | 96.0 – 100.0%  |
|  Systemic sclerosis | 66 | 66 | 100.0% | 94.6 – 100.0%  |
|  Sjögren’s syndrome | 55 | 54 | 98.2% | 90.3 – 100.0%  |
|  MCTD | 45 | 44 | 97.8% | 88.2 – 99.9%  |
|  Other autoimmune diseases* | 63 | 63 | 100.0% | 94.3 – 100.0%  |
|  Borreliosis(5) | 30 | 30 | 100.0% | 88.4 – 100.0%  |
|  Total | 500 | 496 | 99.2% | 98.0 – 99.8%  |

* Samples include: autoimmune hepatitis (n=8), primary biliary cirrhosis (n=9), Grave’s disease (n=12), Hashimoto (n=11), celiac disease (n=11), Diabetes Type I (n=12)

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

Not applicable

4. Clinical cut-off:

See assay cut-off

5. Expected values/Reference range:

The levels of anti-nRNP/Sm, anti-Sm, anti-SS-A, anti-SS-B, anti-Scl-70, anti-Centromeres, anti-Jo-1 in healthy people were investigated with a total of 200 samples from apparently healthy blood donors (including 120 men and 80 women with an average age of 40 years old; age range from 19 to 68 years). For anti-ribosomal P-proteins, 150 healthy blood donor samples were tested. The samples included 79 mean and 71 women with an average age of 38 and age range from 18 to 67. The results are summarized in the following table:

{19}

|   | Anti-nRNP/Sm | Anti-Sm | Anti-SS-A | Anti-SS-B | Anti-Scl-70 | Anti-Centro. | Anti-Jo-1 | Anti-rib.P-proteins  |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
|  n | 200 | 200 | 200 | 200 | 200 | 200 | 200 | 150  |
|  Positives | 1 | 0 | 2 | 0 | 0 | 1 | 0 | 0  |
|  Negatives | 199 | 200 | 198 | 200 | 200 | 199 | 200 | 150  |
|  prevalence | 0.5% | 0.0% | 1.0% | 0.0% | 0.0% | 0.5% | 0.0% | 0.0%  |
|  Ratio Range | 0.1-1.3 | 0.1-0.3 | 0.0-4.4 | 0.0-0.2 | 0.0-0.1 | 0.0-3.0 | 0.0-0.2 | 0.0-0.7  |
|  Mean | 0.1 | 0.1 | 0.1 | 0.0 | 0.0 | 0.1 | 0.1 | 0.1  |
|  Std | 0.09 | 0.02 | 0.33 | 0.02 | 0.01 | 0.21 | 0.03 | 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.

---

**Source:** [https://fda.innolitics.com/submissions/IM/subpart-f%E2%80%94immunological-test-systems/LKO/K123261](https://fda.innolitics.com/submissions/IM/subpart-f%E2%80%94immunological-test-systems/LKO/K123261)

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