VIRGO AMA ELISA KIT

K971909 · Hemagen Diagnostics, Inc. · DBM · Jun 20, 1997 · Immunology

Device Facts

• Record ID: K971909
• Device Name: VIRGO AMA ELISA KIT
• Applicant: Hemagen Diagnostics, Inc.
• Product Code: DBM · Immunology
• Decision Date: Jun 20, 1997
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.5090
• Device Class: Class 2

Intended Use

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

Device Story

VIRGO AMA ELISA KIT detects antimitochondrial antibodies in human serum; utilizes ELISA methodology. Purified mitochondrial antigen immobilized on microwell plates; patient serum incubated; antibodies bind to antigen. Horseradish peroxidase (HRP)-conjugated secondary antibody binds to patient antibodies; TMB substrate added; HRP catalyzes color change. Color intensity measured via EIA plate reader; optical density indicates antibody presence. Used in clinical laboratories; operated by laboratory technicians. Output provides quantitative/semi-quantitative antibody levels; assists clinicians in diagnosing primary biliary cirrhosis (PBC) when combined with clinical findings and other tests.

Clinical Evidence

Comparative study of 154 serum specimens (68 with liver disease, 86 healthy donors) tested by both the proposed ELISA and the predicate fluorescent assay. Results: 100% relative sensitivity (61/61) for liver disease samples (94.1-100% CI) and 100% agreement for normal samples. Precision studies (inter-assay and intra-assay) performed on 12 samples; %CVs reported for OD and units. Interference testing confirmed no significant effect (<15% variation) from hemoglobin (<500 mg/dL), bilirubin (≤20 mg/dL), or lipids (<3000 mg/dL).

Technological Characteristics

Enzyme-linked immunosorbent assay (ELISA). Components: microwell plates with immobilized purified mitochondrial antigen, HRP-conjugated secondary antibody, TMB substrate. Detection principle: optical density measurement via EIA plate reader. Cutoff established for positive/negative classification. Standalone diagnostic kit.

Indications for Use

Indicated for the detection and measurement of antimitochondrial antibodies in human serum to aid in the diagnosis of primary biliary cirrhosis (PBC) in conjunction with clinical observations and other serological tests.

Regulatory Classification

Identification

An antimitochondrial antibody immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the antimitochondrial antibodies in human serum. The measurements aid in the diagnosis of diseases that produce a spectrum of autoantibodies (antibodies produced against the body's own tissue), such as primary biliary cirrhosis (degeneration of liver tissue) and chronic active hepatitis (inflammation of the liver).

Predicate Devices

• Scimedx Mitochondrial Antibody Test System (000001)

Submission Summary (Full Text)

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K971909

## JUN 20 1997

## 510(k) Summary

#### Submitter's Name/Contact Person 1.

Joseph M. Califano, Regulatory Affairs Manager

### Address

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

[phone] Phone: [phone] Fax: email: [e-mail]

#### Date Prepared

19 May 1997

#### 2. Device Names

Trade Name: VIRGO ® AMA ELISA KIT Common Name: Antimitochondrial antibody Kit (EIA method) Classification Name: Antimitochondrial antibody immunological test system

#### 3. Predicate Device

1

Scimedx Mitochondrial Antibody Test System.

000001

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## 3. Description of Device

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

The ELISA methodology is commonly used for serum antibody evaluations. Purified mitochondrial antigen 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 antibodies remaining after the wash step. In the wells where the second antibody remains bound, the conjugated HRP catalyzes a color change in the substrate, tetramethyl benzidine (TMB). 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 antibodies to mitochondrial antigen in human serum.

### 5.(A) Technological Characteristics

#### Proposed Device

The VIRGO ® AMA ELISA KIT is an enzyme-linked immunosorbent assay. The device utilizes optical density as a measure of antibody presence, with an established cutoff between a positive and a negative reaction.

### Predicate Device

2

The Scimedx Mitochondrial Antibody Test System 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

### Precision

To evaluate precision, both inter-assay and intra-assay studies were conducted. The results are summarized below:

#### A Inter-assay

Twelve different samples {9 positive and 3 negative} were assayed in triplicate twice a day for five different days.

| Sample | Mean OD | Std. Dev. | % CV | Mean Units | Std. Dev. | % CV |
|--------|---------|-----------|------|------------|-----------|------|
| 1 | 1.721 | 0.102 | 5.9 | 8.5 | 0.8 | 9.9 |
| 2 | 1.714 | 0.111 | 6.5 | 8.4 | 0.9 | 10.6 |
| 3 | 1.574 | 0.093 | 5.9 | 7.8 | 0.9 | 11.4 |
| 4 | 1.116 | 0.092 | 8.2 | 5.5 | 0.6 | 11.4 |
| 5 | 0.939 | 0.083 | 8.8 | 4.6 | 0.7 | 15.9 |
| 6 | 0.858 | 0.081 | 9.5 | 4.2 | 0.6 | 13.6 |
| 7 | 0.729 | 0.045 | 6.2 | 3.6 | 0.4 | 12.2 |
| 8 | 0.679 | 0.033 | 4.9 | 3.3 | 0.3 | 10.1 |
| 9 | 0.673 | 0.074 | 11.0 | 3.3 | 0.3 | 10.1 |
| 10 | 0.020 | 0.019 | N/A | 0.1 | 0.1 | N/A |
| 11 | 0.025 | 0.025 | N/A | 0.1 | 0.1 | N/A |
| 12 | 0.027 | 0.015 | N/A | 0.1 | 0.1 | N/A |

The assay controls {Positive, Negative, & Cutoff Serum} were assayed concurrently in triplicate twice each day for each of the five days.

| Sample | Mean OD | Std. Dev. | % CV |
|------------------|---------|-----------|------|
| Negative Control | 0.003 | 0.003 | N/A |
| Positive Control | 1.202 | 0.110 | 9.2 |
| Cutoff Serum | 0.205 | 0.024 | 11.6 |

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# Precision

#### Intra-assay B.

4

The same twelve samples {9 positive and 3 negative} were assayed 20 times in a single run.

| Sample | Mean OD | Std. Dev. | % CV | Mean Units | Std. Dev. | % CV |
|--------|---------|-----------|------|------------|-----------|------|
| 1 | 1.362 | 0.050 | 3.7 | 9.3 | 1.0 | 10.7 |
| 2 | 1.243 | 0.034 | 2.7 | 8.4 | 0.8 | 9.6 |
| 3 | 1.354 | 0.066 | 4.9 | 9.2 | 0.8 | 9.1 |
| 4 | 0.822 | 0.058 | 7.0 | 5.6 | 0.5 | 9.2 |
| 5 | 0.608 | 0.043 | 7.2 | 4.1 | 0.4 | 10.2 |
| 6 | 0.636 | 0.054 | 8.5 | 4.3 | 0.6 | 13.7 |
| 7 | 0.600 | 0.039 | 6.5 | 4.1 | 0.4 | 11.0 |
| 8 | 0.559 | 0.030 | 5.4 | 3.8 | 0.4 | 9.6 |
| 9 | 0.532 | 0.033 | 6.1 | 3.6 | 0.4 | 9.7 |
| 10 | 0.010 | 0.002 | N/A | 0.07 | 0.01 | N/A |
| 11 | 0.009 | 0.002 | N/A | 0.06 | 0.01 | N/A |
| 12 | 0.010 | 0.001 | N/A | 0.07 | 0.01 | N/A |

The assay controls {Positive and Cutoff Serum} were assayed concurrently 20 times.

| Sample | Mean OD | Std. Dev. | % CV |
|------------------|---------|-----------|------|
| Positive Control | 1.047 | 0.040 | 3.8 |
| Cutoff Serum | 0.149 | 0.015 | 10.2 |

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# Comparison Testing

Companson 1 estima
A total of 154 serum specimently assayed by both the production of the predicate
A total of 154 serum specitly books, were concurrently assayed by bellevi A total of 154 serum speciment (68 from individuals with known of the discussion,
86 from normal apparently healthy donos) were concurrently assayed by both the follow: A total 01 104 Solan of 104 series are presented in the tables that he mail more.
device and the proposed device. The results are presented in the tables that more.

# Table 1 Combined known or suspected liver disease specimens. n = 68

| | Predicate Device | | |
|-----------------|------------------|----------|-------|
| | Positive | Negative | Total |
| Proposed Device | | | |
| Positive | 61 | 0 | 61 |
| Negative | 0 | 7 | 7 |
| Total | 61 | 7 | 68 |

relative Sensitivity = 100.0 % {61/61}, ... confidence interval = 94.1% to 100 %
Relative Sensitivity = 100.0 % {61/61}, ... confidence interval = 94.1% to 100 %
{{ if(

# Table 2 Normals. n = 86

# Predicate Device

| | Positive | Negative | Total |
|-----------------------------|----------|----------|-------|
| Proposed Device<br>Positive | 0 | 0 | 0 |
| Negative | 0 | 86 | 86 |
| Total | 0 | 86 | 86 |

. ---------------------------------------------------------------------------------------------------------------------------------------------------------------------------

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## Interfering Substances

Lipemic, icteric, and hemolytic samples were evaluated with the assay following Elpenine, and nemoryto bampio "interference Testing in Clinical NGCCS Document E. The results indicate that there is no significant effect (<15 % variation) on the assay for samples with:

Hemoglobin concentration: Bilirubin concentration: Lipid concentration:

< 500 mg/dL ≤ 20 mg/dL < 3000 mg/dL

#### Prozone

The VIRGO ® AMA ELISA Kit was used to assay several high titered serum samples 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.

#### Conclusions

The results of the comparative studies support the claim that the proposed device is substantially equivalent to the predicate device and performs as an effective screening assay.

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Image /page/6/Picture/1 description: The image shows the seal for the Department of Health & Human Services USA. The seal is circular, with the words "DEPARTMENT OF HEALTH & HUMAN SERVICES USA" arranged around the perimeter. In the center of the seal is a stylized image of three human profiles facing right, with flowing lines beneath them.

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

JUN 20 1007

Joseph M. Califano Manager, Regulatory Affairs HEMAGEN DIAGNOSTICS, INC. 34-40 Bear Hill Road Waltham, MA 02154

乐

Re: K971909 Name: VIRGO® AMA ELISA KIT Regulatory Class: II Product Code: 82 DBM Dated: May 19, 1997 Received: May 23, 1997

Dear Mr. Califano:

We have reviewed your Section 510(k) notification of intent to market the device referenced above and we have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to 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). 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.

If your device is classified (see above) into either class II (Special Controls) or class III (Premarket Approval), it may be subject to such additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 895. A substantially equivalent determination assumes compliance with the current Good Manufacturing Practice requirement, as set forth in the Quality System Regulation (QS) for Medical Devices: General regulation (21 CFR Part 820) and that, through periodic (QS) inspections, the Food and Drug Administration (FDA) will verify such assumptions. Failure to comply with the GMP regulation may result in regulatory action. In addition, FDA may publish further announcements concerning your device in the Federal Register. Please note: this response to your premarket notification submission does not affect any obligation you might have under sections 531 through 542 of the Act for devices under the Electronic Product Radiation Control provisions, or other Federal Laws or Regulations.

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Page 2 -

Under the Clinical Laboratory Improvement Amendments of 1988 (CLIA-88), this device may require a CLIA.complexity.categorization. To.determine if it does, you should. - - contact the Centers for Disease Control and Prevention (CDC) at [phone].

This letter will allow you to begin marketing your device as described in your 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801 and additionally 809.10 for in vitro diagnostic devices), please contact the Office of Compliance at [phone]. Additionally, for questions on the promotion and advertising of your device, please contact the Office of Compliance at [phone]. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers Assistance at its toll free number [phone] or at [phone] or at its internet address "http://www.fda.gov/cdrh/dsmamain.html"

Sincerely yours,

Steven Autman

Steven I. Gutman, M.D., M.B.A. Director Division of Clinical Laboratory Devices Office of Device Evaluation Center for Devices and Radiological Health

Enclosure

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( = HOMAGEN DI AGNUS ( LES ID:6178963748 rage 2/2 Device Name: VIRGO ® AMA ELISA KIT Indication(s) For Use This enzyme-linked immunosorbent assay (ELISA) is indicated for the detection and measurement of antimitochondrial antibodies in human serum. The presence of mitochondrial antibodies, in combination with clinical observations and other serological tests, can aid in the diagnosis of primary biliary cirrhosis(PBC) on of Clinical Laboratory Der (PLEASE DO NOT WRITE BELOW THIS LINE) Concurrence of CDRH, Office of Device Evaluation (ODE) rescription Use OR Over-The-Counter-Use Per 21 CFR 801.109)