EPSTEIN-BARR VIRUS EARLY ANTIGEN (EBV-EA) IGG IFA TEST SYSTEM

K992587 · Stellar Bio Systems, Inc. · LSE · Dec 22, 1999 · Microbiology

Device Facts

• Record ID: K992587
• Device Name: EPSTEIN-BARR VIRUS EARLY ANTIGEN (EBV-EA) IGG IFA TEST SYSTEM
• Applicant: Stellar Bio Systems, Inc.
• Product Code: LSE · Microbiology
• Decision Date: Dec 22, 1999
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3235
• Device Class: Class 1
• Attributes: Real-World Evidence

Real-World Evidence

Submission	Device	Sponsor	RWD Sources	RWE Use Summary	Key Tags
K992587 · Dec 22, 1999	EPSTEIN-BARR VIRUS EARLY ANTIGEN (EBV-EA) IGG IFA TEST SYSTEM	Stellar Bio Systems, Inc.	Retrospective clinical serum samples from a commercial clinical laboratory	Retrospective clinical sera were used to determine the relative sensitivity and specificity of the Stellar EBV-EA IFA kit compared to a commercially available predicate IFA device.	Retrospective clinical sera; Relative sensitivity; Relative specificity; Clinical laboratory samples

Clinical Evidence

Study Design	Population	Comparator	Key Endpoints
Retrospective comparative study	Human serum samples (acute, seronegative, and seropositive); Sample Size: 75; Number of Sites: 1	Commercially available EBV-EA IFA	Relative sensitivity and specificity

Indications for Use

The Stellar Bio Systems' Indirect Fluorescence Assay (IFA) for Epstein-Barr Early Antigen (EBV-EA) IgG Antibody is intended for the qualitative and semi-quantitative detection of IgG (Immunoglobulin G) antibody to the early antigen complex of the Epstein-Barr Virus in human serum. The test detects IgG antibodies to both the diffuse (D) and restricted (R) components of the EBV early antigen complex. The EBV-EA IFA Test Kit should be used in combination with other Epstein-Barr serologies (Viral Capsid Antigen (VCA) IgG and IgM, Epstein-Barr Nuclear Antigen (EBNA), and heterophile antibody) as an aid in the diagnosis of infectious mononucleosis (IM).

Device Story

In vitro diagnostic test; detects IgG antibodies to Epstein-Barr Virus Early Antigen (EBV-EA) in human serum. Principle: Indirect Fluorescence Assay (IFA). Patient serum applied to fixed antigens on glass microscope slides; incubated; washed; fluorescein-conjugated goat anti-human IgG applied; incubated; washed. Results visualized via fluorescence microscopy; positive reaction indicated by bright green fluorescence at antigen sites. Used in clinical laboratories by trained personnel. Output: qualitative or semi-quantitative titer. Assists clinicians in diagnosing infectious mononucleosis when used alongside other EBV serological markers.

Clinical Evidence

Bench testing only. Evaluated using 75 retrospective frozen human sera samples compared to a commercial IFA. Overall agreement was 90.7% (95% CI: 81.7% - 96.2%). Titer agreement study (n=25) showed 9/25 identical titers and 16/25 within one two-fold dilution. Reproducibility study (n=180) showed 131/180 identical titers and 49/180 within one two-fold dilution. Specificity testing (n=10) showed no cross-reactivity with other herpesviruses (CMV, VZV, HSV, HHV).

Technological Characteristics

Indirect Fluorescence Assay (IFA). Components: chemically induced Raji cells (antigen source), fluorescein-conjugated goat anti-human IgG. Form factor: paint-delineated wells on glass microscope slides. Detection: fluorescence microscopy. No electronic or software components.

Indications for Use

Indicated for qualitative and semi-quantitative detection of IgG antibodies to EBV early antigen complex (diffuse and restricted components) in human serum. Used as an aid in the diagnosis of infectious mononucleosis (IM) in conjunction with other EBV serologies (VCA IgG/IgM, EBNA, heterophile antibody).

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).

Submission Summary (Full Text)

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# DEC 2 2 1999

K992587

# 510k 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:

### Applicant Information:

| Date Prepared: | July 20, 1999 |
|-----------------|------------------------------------------|
| Name: | Stellar Bio Systems, Inc |
| Address: | 9075 Guilford Road<br>Columbia, MD 21046 |
| Contact Person: | John Brewer |

| Contact Person: | John Brewer |
|-----------------|--------------|
| PhoneNumber. | 410-381-8550 |
| Fax Number. | 410-381-8984 |

#### Device Information:

| Trade Name: | Indirect Fluorescence Assay for Epstein-Barr Virus Early Antigen (EBV-EA) IgG<br>Antibody |
|----------------------|-------------------------------------------------------------------------------------------|
| Common Name: | EBV-EA IgG IFA Test |
| Classification Name: | EBV-EA IgG Serological Reagent |

### Equivalent Device: IFA

Device Description: The Epstein-Barr Virus Early Antigen (EBV-EA) IgG IFA is an Indirect Fluorescence Assay (IFA) for the detection of IgG antibodies to EBV-EA in human serum.

Intended Use: The Stellar Bio Systems' Indirect Fluorescence Assay (IFA) for Epstein-Barr Virus Early Antigen (EBV-EA) IgG Antibody is intended for the qualitative and semi-quantitative detection of IgG (Immunoglobulin G) antibody to the early antigen complex of the Epstein-Barr Virus in human serum. The test detects IgG antibodies to EBV antigens in chemically induced Raji cells. The EBV-EA IFA Test Kit should be used in combination with other Epstein-Barr serologies [Viral Capsid Antigen (VCA) IgG and IgM, Epstein-Barr Nuclear Antigen (EBNA), and heterophile antibody] as an aid in the diagnosis of infectious mononucleosis (IM).

### Principle of Procedure:

Stellar Bio Systems' fluorescent antibody assays use the indirect method of antibody detection and titer determination. Diluted patient serum sampled to fixed antigens provided on paint delineated wells on glass microscope slides. During a 30 minute incubation, antibody specific for EA forms an antigen/antibody complex with the fixed antigens. In a brief washing step, nonspecific antibody and other unreacted serum proteins are eliminated. Fluorescein-conjugated goat anti-human IgG is then applied to the glass slide. The anti-IgG conjugate combines with human IgG, if present, during a 30 minute incubation. After a brief wash to remove unreacted conjugate, the slides are viewed by fluorescence microscopy. A positive antibody reaction is denoted by bright green fluorescence at the antigen sites.

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## Performance Characteristics

- 1. Relative Sensitivity and Specificity The Stellar EBV-EA IFA kit was evaluated relative to a commercially available EBV-EA IFA. The samples were selected frozen retrospective sera tested at a commercial clinical laboratory. The acute sera were selected by being positive in a heterophile assay. The seronegative sera were selected by being negative for VCA G and EBNA G by ELISA. The seropositive sera were selected by being positive for VCA G and EBNA G by ELISA.. The data in Table 1 summarizes the data.
# Table 1 Relative Sensitivity and Specificity of the Stellar EBV-EA IFA Kit Relative to Alternate IFA

| | | Stellar EBV-EA IFA | | |
|------------------|----------|--------------------|----------|-------|
| | | Positive | Negative | Total |
| | Positive | 42 | 1 | 43 |
| Alternate<br>IFA | Negative | 6* | 26 | 32 |
| | Total | 48 | 27 | 75 |

95% Confidence Interval Overall Agreement = 68/75 = 90.7% 81.7% - 96.2%

* The 6 Stellar positive/Alternate IFA negative samples were acute sera determined by ELISA VCA IgM.

Please be advised that 'relative' refers to the comparison of this assay's results to that of a similar assay. There was not an attempt to correlate the assay's results with disease presence or absence. No judgment can be made on the comparison assay's accuracy to predict disease.

Since the above studies were performed on a pre-selected retrospective population no calculations for the assay's positive and negative predictive value may be done or inferred.

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The above sera were categorized relative to their serological status to EBV. The following table illustrates the results:

| | Acute<br>VCA IgM (+)<br>EBNA (-) | Seropositive<br>VCA IgG (+)<br>EBNA (+) VCA IgM (-) | Seronegative<br>VCA IgG (-)<br>EBNA (-) |
|-------------------|----------------------------------|-----------------------------------------------------|-----------------------------------------|
| EBV EA<br>IgG IFA | | | |
| Positive | 20 | 25 | 3 |
| Negative | 10 | 7 | 10 |
| Total | 30 | 32 | 13 |

- 2. Titer Agreement: 25 positive sera were serially two-fold diluted and the endpoint titer was determined on the Stellar EBV-EA IFA and a commercially available EBV-EA IFA. The endpoint titer results are as follows.

| Identical Titer | 9/25 |
|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------|
| + one, two-fold dilution<br>- | 12/25 |
| + two, two-fold dilutions<br>------------------------------------------------------------------------------------------------------------------------------------------------------------------------------ | 4/25 |

- 3. Reproducibility: Three positive sera with various titers (1:80,1:160, 1:640) and one negative sera were serially diluted and assayed five times each on three different assays at three different sites. 131/180 of the end point titers were identical. 49/180 of the end point titers was within + one, two-fold dilution.
#### 4. Specificity

The following sera that were positive for IgG antibodies to CMV, VZV, HSV 1, HSV 2, HHV 6 and HHV 8 by IFA were found to be negative for EBV-EA IFA, indicating a lack of cross-reactivity to these viruses.

| Sample | EA IFA Result |
|-----------|---------------|
| CMV + | - |
| VZV + | - |
| HSV 1&2 + | - |
| HSV 1&2 + | - |
| HSV 1&2 + | - |
| HSV 1&2 + | - |
| HHV 6+ | - |
| HHV 6+ | - |
| HHV 8+ | - |

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Image /page/3/Picture/1 description: The image is a black and white logo for the U.S. Department of Health & Human Services. The logo features a stylized image of an eagle with three lines representing its body and wings. The words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" are arranged in a circular pattern around the eagle.

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

DEC 22 1999

Mr. John Brewer President Stellar Bio Systems, Inc. 9075 Guilford Road Columbia, Maryland 21046

Re: K992587

> Trade Name: Indirect Fluorescence Assay (IFA) for Epstein-Barr Virus Early Antigen (EBV-EA) IgG Antibody Regulatory Class: I

:"... Product Code: LSE Dated: October 29, 1999 Received: October 29, 1999

Dear Mr. Brewer:

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 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). 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 requirements, 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 Sutman

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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510(k) Number (if known): K992587

Device Name: Indirect Fluorescence Assay (IFA) for Epstein-Barr Virus Early Antigen (EBV-EA) IgG Antibody

Indications For Use: The Stellar Bio Systems' Indirect Fluorescence Assay (IFA) for Epstein-Barr Early Antigen (EBV-EA) IgG Antibody is intended for the qualitative and semi-quantitative detection of IgG (Immunoglobulin G) antibody to the early antigen complex of the Epstein-Barr Virus in human serum. The test detects IgG antibodies to both the diffuse (D) and restricted (R) components of the EBV early antigen complex. The EBV-EA IFA Test Kit should be used in combination with other Epstein-Barr serologies (Viral Capsid Antigen (VCA) IgG and IgM, Epstein-Barr Nuclear Antigen (EBNA), and heterophile antibody) as an aid in the diagnosis of infectious mononucleosis (IM).

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

Concurrence of CDRH, Office of Device Evaluation (ODE)

Woody Duboe

(Division Sign Off)
Division of Clinical Laboratory Devices
510(k) Number K992587

Prescription Use (Per 21 CFR 801.109)

OR

Over-The-Counter Use

(Optional Format 1-2-96)