EBV V VCA IGM ELISA KIT MODEL# EBM-200

K020717 · Pan Bio Pty. , Ltd. · GNP · Jun 13, 2002 · Microbiology

Device Facts

• Record ID: K020717
• Device Name: EBV V VCA IGM ELISA KIT MODEL# EBM-200
• Applicant: Pan Bio Pty. , Ltd.
• Product Code: GNP · Microbiology
• Decision Date: Jun 13, 2002
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3235
• Device Class: Class 1

Intended Use

The Epstein-Barr Viral Capsid Antigen (VCA) IgM ELISA Test is for the qualitative detection of IgM antibodies to EBV VCA in serum as an aid in the clinical laboratory diagnosis of EBV infection in patients with clinical symptoms consistent with infectious monosis (IM). The PANBIO EBV VCA IgM ELISA should be used in conjunction with other EBV serology.

Device Story

The EBV-VCA IgM ELISA Kit is an in vitro diagnostic assay used in clinical laboratories to detect IgM antibodies to Epstein-Barr Viral Capsid Antigen in human serum. The device utilizes immunopurified EBV-VCA antigen (specific for gp125) coated onto polystyrene microwells. Patient serum is added; if anti-VCA IgM antibodies are present, they bind to the antigen. After washing, peroxidase-conjugated anti-human IgM is added, followed by a TMB/H2O2 substrate system. The enzymatic reaction produces a color change (blue to yellow upon acidification), which is measured to indicate the presence of antibodies. Results are interpreted by laboratory personnel to assist in the diagnosis of EBV infection in patients presenting with symptoms of infectious mononucleosis. The test is intended to be used in conjunction with other EBV serological markers.

Clinical Evidence

Clinical performance was evaluated using retrospective (n=156) and prospective (n=352) serum samples. In the retrospective study, sensitivity for acute IM was 100% (26/26) and specificity for negative samples was 100% (28/28). In the prospective study, sensitivity for acute IM was 85.7% (36/42) and specificity for negative samples was 87.5% (42/48). Analytical specificity was confirmed using a 50-specimen panel of patients with other diseases (CMV, VZV, HSV, ANA, Rheumatoid Factor), showing 0% cross-reactivity. Reproducibility was assessed via ANOVA across three sites with total CVs ranging from 4.8% to 28.1%.

Technological Characteristics

Enzyme-linked immunosorbent assay (ELISA) using immunopurified EBV-VCA antigen (gp125) immobilized on polystyrene microwells. Detection utilizes peroxidase-conjugated anti-human IgM and TMB/H2O2 substrate system. Manual or automated plate processing. No software algorithm or electronic connectivity described.

Indications for Use

Indicated for qualitative detection of IgM antibodies to EBV-VCA in human serum as an aid in diagnosing EBV infection in patients with clinical symptoms of infectious mononucleosis (IM).

Regulatory Classification

Identification

Epstein-Barr virus serological reagents are devices that consist of antigens and antisera used in serological tests to identify antibodies to Epstein-Barr virus in serum. The identification aids in the diagnosis of Epstein-Barr virus infections and provides epidemiological information on diseases caused by these viruses. Epstein-Barr viruses are thought to cause infectious mononucleosis and have been associated with Burkitt's lymphoma (a tumor of the jaw in African children and young adults) and postnasal carcinoma (cancer).

Predicate Devices

• DiaSorin Incstar EBV-VCA IgM ELISA

Submission Summary (Full Text)

{0}------------------------------------------------

# JUN 1 3 2002

# 510(k) SUMMARY OF SAFETY AND EFFECTIVENESS

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.

The assigned 510(k) number is: K020717

## Applicant Information:

| Date Prepared: | 07th June, 2002 |
|----------------|---------------------------------------------|
| Name: | PANBIO Limited |
| Address: | 116 Lutwyche Road<br>Windsor 4030 Australia |

| Contact Person: | Helen Jennings |
|-----------------|-------------------|
| Phone Number. | 61-(0)7-3357-1177 |
| Fax Number. | 61-(0)7-3357-1222 |

#### Device Information:

| Trade Name: | EBV-VCA IgM ELISA Kit |
|----------------------|---------------------------------|
| Common Name. | EBV-VCA IgM EIA Test |
| Classification Name; | EBV-VCA IgM Serological Reagent |

#### Equivalent Device:

DiaSorin Incstar EBV-VCA IgM ELISA

#### Device Description:

The EBV-VCA IgM ELISA Kit is an enzyme-linked immunosorbent assay (ELISA) for the detection of IgM antibodies to EBV-VCA antigen in human serum.

## Intended Use:

The Epstein-Barr Viral Capsid Antigen (VCA) IgM ELISA Test is for the qualitative detection of IgM antibodies to EBV VCA in serum as an aid in the clinical laboratory diagnosis of EBV infection in patients with clinical symptoms consistent with infectious mononucleosis (IM). The PANBIO EBV VCA IgM ELISA should be used in conjunction with other EBV serology.

#### Principle of Procedure:

Serum containing antibodies to VCA antigen, when present, combine with EBV-VCA antigen, which is immunopurified utilising a monoclonal specific for gp125 and attached to the polystyrene surface of the microwells. Residual serum is removed by washing and peroxidase conjugated anti-human IgM is added. The microwells are washed and a colourless substrate system, tetramethylbenzidine/ hydrogen peroxide (TMB/H2/O2) is added. The substrate is hydrolysed by the enzyme and the chromogen changes to a blue colour. After stopping the reaction with acid, the TMB becomes yellow. Color development is indicative of the presence of EBV-VCA IgM antibodies in the test sample.

{1}------------------------------------------------

## PERFORMANCE CHARACTERISTICS

#### Study Site 2:

156 frozen retrospective sera of various ages and genders were submitted to a state health lab in Maryland USA for EBV testing. The sera include samples from the following groups: 28 seronegative samples, 26 samples from patients with acute Infectious Mononucleosis, and 102 samples from patients with past exposure to EBV. These sera were tested on the PANBIO EBV-VCA IgM kit and EBV ELISA assays from an alternate manufacturer to determine the EBV status of the sera. The PANBIO EBV VCA IgM results were compared to the EBV status of the sera to determine the sensitivity, specificity, and agreement of the assay relative to the EBV serological status. The data is summarized in the Table 1.

# TABLE 1 EBV VCA IgM Serological Sensitivity and Specificity of PANBIO ELISA versus EBV Status

| EBV Status | Positive | Equivocal* | Negative | Total |
|--------------------------------------------------------------|----------|------------|----------|-------|
| Seronegative<br>VCA IgG (-)<br>VCA IgM (-)<br>EBNA IgG (-) | 0 | 0 | 28 | 28 |
| Acute<br>VCA IgM (+)<br>EBNA IgG (-) | 26 | 0 | 0 | 26 |
| Past Infection<br>VCA IgG (+)<br>VCA IgM (-)<br>EBNA IgG (+) | 5 | 5 | 92 | 102 |
| Total | 31 | 5 | 120 | 156 |

PANBIO ELISA

#### 95% Confidence Interval

| Serological Sensitivity (Acute) | = 26/26 | = 100 % | 86.8 – 100 % |
|------------------------------------|-----------|----------|---------------|
| Serological Sensitivity (Past) | = 92/102 | = 90.2 % | 82.7 – 95.2 % |
| Serological Specificity (Negative) | = 28/28 | = 100% | 87.7 – 100 % |
| Serological Agreement | = 146/156 | = 93.6 % | 88.5 – 96.9 % |

* Retesting of equivocal samples was not conducted, as the samples were unavailable.

Note: "Serological" sensitivity and specificity refers to the comparison of the PANBIO assay results to other assays normally used to diagnose EBV associated IM. There was not an attempt to correlate the assay's results with disease presence or absence. No judgement can be made on the comparison's accuracy to predict disease. Since the above studies were performed on a pre-selected, retrospective, populations for the assay's positive and negative predictive value may be done or inferred.

{2}------------------------------------------------

## Study Site 3:

352 prospective sera of various ages and genders were submitted to a private pathology laboratory in Oueensland Australia for EBV testing. The sera include samples from the following groups: 48 seronegative samples, 42 samples from patients with acute Infectious Mononucleosis, and 262 samples from patients with past exposure to EBV. These sera were tested on the PANBIO EBV-VCA IgM kit and EBV ELISA assays from an alternate manufacturer to determine the EBV status of the PANBIO VCA IgM results were compared to the EBV status of the sera to determine the sensitivity, specificity, and agreement of the assay relative to the EBV serological status. The data is summarized in Table 2.

#### PANBIO ELISA EBV Status Positive Equivocal* Negative Total Seronegative VCA IgG (-) 5 l 42 48 VCA IgM (-) EBNA IgG (-) Acute 36 5 VCA IgM (+) 1 42 EBNA IgG (-) Past Infection VCA IgG (+) 5 0 257 262 VCA IgM (-) EBNA IgG (+) 2 Total 46 304 352 95% Confidence Interval Serological Sensitivity (Acute) = 36/42 = 85.7% 71.5 - 94.6% Serological Specificity (Past) = 257/262 =98.1% 95.6-99.4% Serological Specificity (Negative) = 42/48 74.7 - 95.3% = 87.5%

**PANBIO ELISA**

TABLE 2 EBV-VCA IgM Serological Sensitivity and Specificity of PANBIO ELISA versus EBV Status

* These equivocal samples were not tested on an alternative method due to insufficient sample. Collection of a further sample was not possible.

= 95.2%

92.4 - 97.2%

= 335/352

Serological Agreement

{3}------------------------------------------------

# REPRODUCIBILITY

The reproducibility of the PANBIO EBV-VCA IgM ELISA kit was determined by testing 8 sera 3 times each on three different days at three Australian study sites. Two sites were private pathology laboratories and the third site was PANBIO. Within-run, between day, between site and total precision were estimated by analysis of variance (ANOVA Type II) and are presented in table 3 below.

# TABLE 3 REPRODUCIBILITY DATA PANBIO EBV-VCA IgM Study Site 1, 2 & 3 Precision Measures (Using Cut-Off Ratio)

| | | Within | | Between Day | | Between Site | | Total | | |
|----------|----|--------|------|-------------|------|--------------|------|-------|------|-------|
| Sample | n | *Mean | *S.D | CV | *S.D | CV | *S.D | CV | *S.D | CV |
| Positive | 27 | 2.38 | 0.17 | 6.9% | 0.09 | 3.9% | 0.01 | 0.4% | 0.18 | 7.7% |
| Cut-off | 27 | 1.00 | 0.05 | 5.2% | 0.00 | 0.0% | 0.00 | 0.0% | 0.05 | 4.8% |
| Negative | 27 | 0.10 | 0.02 | 15.5% | 0.02 | 15.5% | 0.01 | 9.4% | 0.02 | 21.6% |
| #1 | 27 | 2.97 | 0.19 | 6.3% | 0.23 | 7.8% | 0.13 | 4.2% | 0.29 | 9.7% |
| #2 | 27 | 3.20 | 0.19 | 5.9% | 0.19 | 6.0% | 0.06 | 1.9% | 0.25 | 7.9% |
| #3 | 27 | 1.20 | 0.13 | 10.5% | 0.13 | 10.9% | 0.00 | 0.0% | 0.16 | 13.6% |
| #4 | 27 | 1.28 | 0.06 | 5.0% | 0.05 | 3.9% | 0.06 | 4.8% | 0.09 | 7.2% |
| #5 | 27 | 0.65 | 0.12 | 18.3% | 0.05 | 8.1% | 0.01 | 1.5% | 0.13 | 19.5% |
| #6 | 27 | 0.95 | 0.08 | 8.8% | 0.06 | 6.3% | 0.00 | 0.0% | 0.10 | 10.1% |
| #7 | 27 | 3.45 | 0.24 | 7.1% | 0.32 | 9.1% | 0.00 | 0.0% | 0.35 | 10.2% |
| #8 | 27 | 1.37 | 0.27 | 19.8% | 0.31 | 22.8% | 0.10 | 7.4% | 0.38 | 28.1% |

All values are calculated from Ratios (Cut-Off using O.D) SD = Standard Deviation; CV = Coefficient of Variation

Note: Standard Deviation results have been rounded to two decimal places for tabulation purposes

{4}------------------------------------------------

## POTENTIAL CROSS-REACTIVITY

## Study Site 5:

A panel of 50 specimens from patients with confirmed diseases other than Enstein Barr Virus was tested to establish the analytical specificity of the EBV-VCA IgM ELISA Test. The specimens were from patients with diseases that have the potential for cross-reactivity. Each of the specimens included in the study was characterized with respect to disease diagnosis prior to analysis with the EBV-VCA IgM ELISA Test. Table 4 lists a summary of the results.

## TABLE 4

| Disease Type | Number of Specimens | Positive Result |
|----------------------------|---------------------|-----------------|
| Cytomegalovirus | 9 | (0/9) |
| Varicella zoster | 10 | (0/10) |
| Herpes simplex virus 1 & 2 | 8 | (0/8) |
| Anti-Nuclear Antigen | 14 | (0/14) |
| Rheumatoid Factor | 9 | (0/9) |
| Total | 50 | (0/50) |

## PANBIO EBV-VCA IgM CROSS-REACTIVITY SPECIMEN PANEL

Results indicate that no specimens (0/50) were positive when analysed with the EBV-VCA IgM ELISA Kit. Refer to 'Study Document -- Site 5' for raw data and section 2.3.4.1 for the summary table.

The true negative result of 100% for the above disease panel is consistent with good analytical specificity for the EBV-VCA IgM ELISA Test.

{5}------------------------------------------------

Image /page/5/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES USA" around the perimeter. Inside the circle is a stylized image of three overlapping human profiles facing to the right, with three curved lines above them, resembling wings or feathers.

JUN 1 3 2002

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Ms. Helen Jennings Ouality and Regulatory Affairs Manager PANBIO Limited 116 Lutwyche Road Windsor, Brisbane Queensland, 4030 Australia

Re: k020717

> Trade/Device Name: EBV Viral Capsid Antigen IgM ELISA Test Regulation Number: 21 CFR 866.3235 Regulation Name: Epstein - Barr virus Serological Reagents Regulatory Class: Class I Product Code: GNP Dated: May 9, 2002 Received: May 15, 2002

Dear Ms. Jennings:

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. Iisting 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 (PMA), 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 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801); 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.

{6}------------------------------------------------

Page 2 -

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 1 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" (21CFR 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers International and Consumer Assistance at its toll-free number [phone] or [phone] or at its internet address "http://www.fda.gov/cdrh/dsma/dsmamain.html".

Sincerely yours,

Steven Gutman

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

{7}------------------------------------------------

Page 1 of of 1

# 510(k) Number (if known): K020717

#### EBV-VCA IgM ELISA Device Name:

Indications For Use: The Epstein-Barr Viral Capsid Antigen (VCA) IgM ELISA Test is for the qualitative detection of IgM antibodies to EBV VCA in serum as an aid in the clinical laboratory diagnosis of EBV infection in patients with clinical symptoms consistent with infectious mononucleosis (IM). The PANBIO EBV VCA IgM ELISA should be used in conjunction with other EBV serology.

(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH, Office of Device Evaluation (ODE)

Woody Dubois

Chnical Laboratory Devices K020 717 510(k) Number .

PRESCRIPTION USE X

(Optional Format 3-10-98)