← Product Code [DHN](/productcode/DHN) · K962276

# HEMAGEN ENA DSDNA KIT (EIA METHOD) (K962276)

_Hemagen Diagnostics, Inc. · DHN · Dec 5, 1996 · Immunology · SESE_

**Canonical URL:** https://fda-staging.innolitics.com/device/K962276

## Device Facts

- **Applicant:** Hemagen Diagnostics, Inc.
- **Product Code:** [DHN](/productcode/DHN.md)
- **Decision Date:** Dec 5, 1996
- **Decision:** SESE
- **Submission Type:** Traditional
- **Regulation:** 21 CFR 866.5100
- **Device Class:** Class 2
- **Review Panel:** Immunology

## Indications for Use

An enzyme-linked immunosorbent assay (ELISA) designed for the detection and measurement of circulating antibodies to native DNA. These autoantibodies are often associated with systemic lupus erythematosus (SLE) and give positive results in screening tests for antinuclear antibodies.

## Device Story

Hemagen DNA Kit is an ELISA for detecting/measuring anti-native DNA antibodies in human serum. Purified native DNA is immobilized on microwell plates. Patient serum is incubated; specific antibodies bind to antigen. HRP-conjugated secondary antibody binds to patient DNA antibodies; HRP catalyzes color change in substrate. Color intensity measured via EIA plate reader. Used in clinical laboratory settings. Output is optical density, correlated to antibody presence/concentration. Assists clinicians in screening for antinuclear antibodies associated with systemic lupus erythematosus (SLE).

## Clinical Evidence

Bench testing and comparative clinical study. Precision evaluated via inter-assay (50 readings) and intra-assay (20 consecutive runs) studies. Comparative study of 140 samples (100 autoimmune, 40 healthy) against predicate IFA kit. Results: 100% relative analytical sensitivity (23/23) and 97.4% relative analytical specificity (75/77). Interference testing performed for hemoglobin (≤500 mg/dL), lipids (≤3000 mg/dL), and bilirubin (≤20 mg/dL) per NCCLS EP7-P, showing <15% variation. Prozone effect evaluated with high-titer samples, showing no false-low results.

## Technological Characteristics

ELISA-based immunoassay. Purified native DNA immobilized on microwell plates. HRP-conjugated secondary antibody detection system. Optical density measurement via EIA plate reader. Calibrated against WHO Standard for Anti-Double Stranded DNA (Code Wo/80).

## 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

- Hemagen VIRGO Anti-nDNA IgG IFA Kit (K771376A)

## Submission Summary (Full Text)

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K962276

510(k) Summary

1.  Submitter's Name/Contact Person
DEC - 5 1996
Joseph M. Califano, Manager, Regulatory Affairs

Address
Hemagen Diagnostics, Inc.
34-40 Bear Hill Road
Waltham, MA, 02154

Phone: (617) 890-3766
Fax: (617) 890-3748

Date Prepared: 14 June 1996

2.  Device Name

Trade Name: Hemagen ® DNA Kit (EIA method)
Common Name: Anti-DNA
Classification Name: Anti-DNA Antibody, Antigen and Control

3.  Predicate Device

Hemagen VIRGO ® Anti-nDNA IgG IFA Kit
{Reference 510 (k) No. K 771376A}

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## 3. Description of Device

An enzyme-linked immunosorbent assay (ELISA) designed for the detection and measurement of antibodies to native DNA in human serum.

The ELISA methodology is commonly used for serum antibody evaluations. Purified native DNA has been attached to the inner surfaces of the microwell plate. During the initial incubation step, antibodies in patient serum bind specifically to the immobilized antigen and remain in place after a wash step.

A second antibody which is conjugated to horseradish peroxidase (HRP) is used to recognize the "heavy + light" chain regions of the patient's DNA antibodies that remain after the wash step. In the wells where the second antibody remains bound, the conjugated HRP catalyzes a color change in the substrate. After the reaction is stopped, the color is read in an EIA Plate reader.

## 4. Intended Use of Device

An enzyme-linked immunosorbent assay (ELISA) designed for the detection and measurement of circulating antibodies to native DNA. These autoantibodies are often associated with systemic lupus erythematosus (SLE) and give positive results in screening tests for antinuclear antibodies.

### 5.(A) Technological Characteristics

#### Proposed Device

The Hemagen dsDNA Kit is an enzyme-linked immunosorbent assay. The device utilizes optical density as a measure of antibody presence, with an established cutoff point between a positive and a negative reaction.

#### Predicate Device

The Hemagen VIRGO Anti-DNA IgG {heavy + light} IFA Kit is an indirect fluorescent antibody assay. The device utilizes the indirect method of fluorescent antibody staining. The resultant level of observed fluorescence is used to determine the presence or absence of antibodies.

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# 5.(B) Performance Data

## I. Precision

To evaluate precision, inter-assay and intra-assay studies were conducted.

### A. Inter-assay reproducibility {Between-run}

Eight different serum samples were assayed five times each, twice a day, on five different days (a total of 50 readings)

|  Sample | Mean IU/mL | Std. Dev. | % CV | Mean O.D. | Std. Dev. | % CV  |
| --- | --- | --- | --- | --- | --- | --- |
|  1 | 102.1 | 11.3 | 11.1 | 0.617 | 0.05 | 8.3  |
|  2 | 102.4 | 6.4 | 6.2 | 0.620 | 0.05 | 7.4  |
|  3 | 90.1 | 6.5 | 7.2 | 0.553 | 0.04 | 7.7  |
|  4 | 55.3 | 5.2 | 9.4 | 0.356 | 0.04 | 9.9  |
|  5 | < 50 | N/A | N/A | 0.213 | 0.03 | 12.4  |
|  6 | < 50 | N/A | N/A | 0.264 | 0.03 | 9.8  |
|  7 | 122.7 | 13.8 | 11.3 | 0.731 | 0.09 | 12.3  |
|  8 | 142.6 | 9.2 | 6.4 | 0.840 | 0.07 | 8.3  |

### B. Intra-assay reproducibility {Within-run}

Eight different samples were assayed 20 consecutive times in a single run:

|  Sample | Mean IU/mL | Std. Dev. | % CV | Mean O.D. | Std. Dev. | % CV  |
| --- | --- | --- | --- | --- | --- | --- |
|  1 | 105.2 | 17.9 | 17.0 | 0.604 | 0.08 | 13.9  |
|  2 | 97.2 | 2.8 | 2.8 | 0.567 | 0.01 | 2.2  |
|  3 | 84.9 | 2.2 | 2.6 | 0.505 | 0.01 | 2.2  |
|  4 | 54.3 | 1.7 | 2.6 | 0.355 | 0.01 | 2.4  |
|  5 | < 50 | N/A | N/A | 0.199 | 0.02 | 9.0  |
|  6 | < 50 | N/A | N/A | 0.240 | 0.01 | 5.5  |
|  7 | 115.4 | 5.6 | 4.8 | 0.688 | 0.03 | 4.2  |
|  8 | 129.4 | 3.8 | 2.9 | 0.765 | 0.02 | 2.6  |

## II. Verification of the DNA Calibrators

The kit calibrators have been compared to the World Health Organization Standard for Anti-Double Stranded DNA (ANTI-dsDNA) 1st International Standard, Code Wo / 80. A study was conducted to demonstrate the high degree of correlation that exists between the kit calibrators and the W.H.O. Standard.

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# III. Comparison Testing

The Hemagen dsDNA Kit and the Hemagen VIRGO Anti-nDNA IgG IFA Kit were used to assay 100 serum specimens from different patients with known or suspected autoimmune disease. Forty (40) specimens from apparently healthy blood donors were also assayed with the Hemagen dsDNA Kit and the Hemagen VIRGO Anti-nDNA IgG IFA Kit.

## Summary Tables

### Table A: Summary of disease state patients, N = 100 (Initial testing)

#### Predicate Device

|   | Positive | Negative | TOTAL  |
| --- | --- | --- | --- |
|  Proposed Device |  |  |   |
|  Positive | 23 | 2^{1} | 25  |
|  Negative | 7^{1} | 68 | 75  |
|  Totals | 30 | 70 | 100  |

1. The nine discrepant samples were evaluated by hemagglutination for resolution

### Table B: Summary of disease state patients, N = 100 (Final results)

#### Predicate Device

|   | Positive | Negative | TOTAL  |
| --- | --- | --- | --- |
|  Proposed Device |  |  |   |
|  Positive | 23 | 2 | 25  |
|  Negative | 0 | 75 | 75  |
|  Totals | 23 | 77 | 100  |

The relative analytical sensitivity was found to be 100 % (23/23).
The relative analytical specificity was found to be 97.4 % (75/77).

### Table C: Normal blood donors, N = 40

#### Predicate Device

|   | Positive | Negative | TOTAL  |
| --- | --- | --- | --- |
|  Proposed Device |  |  |   |
|  Positive | 0 | 0 | 0  |
|  Negative | 0 | 40 | 40  |
|  Totals | 0 | 40 | 40  |

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# IV. Interfering Substances

Lipemic, hemolytic, and icteric samples were evaluated with the assay in accordance with NCCLS Document EP7-P Proposed Guideline "Interference Testing in Clinical Chemistry." The results indicate that there is no significant effect (&lt;15 % variation) on the assay for samples with:

- Hemoglobin concentration: ≤ 500 mg/dL
- Lipid concentration: ≤ 3000 mg/dL
- Bilirubin concentration: ≤ 20 mg/dL

# V. Prozone

The Hemagen dsDNA Kit was used to assay a high titered serum sample to determine if the kit would return unexpectedly low values. The results of this evaluation indicate that the kit gives appropriately high positive results with high titered sera.

# 6. Conclusion

The results of the comparative studies support the claim that the Hemagen dsDNA Kit is substantially equivalent to the predicate device.

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**Source:** [https://fda-staging.innolitics.com/device/K962276](https://fda-staging.innolitics.com/device/K962276)

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