EBV EA-D IGG ELISA KIT, MODEL EBG-100

{0}------------------------------------------------ # SEP 2 7 2002 #### 510(k) SUMMARY OF SAFETY AND EFFECTIVENESS 1.10 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: K021793 ## Applicant Information: | Date Prepared: | September 26, 2002 | |-----------------|---------------------------------------------| | Name: | PANBIO Limited | | Address: | 116 Lutwyche Road<br>Windsor 4030 Australia | | Contact Person: | Helen Jennings | | Phone Number: | 61-(0)7-3357-1177 | ## Device Information: Fax Number: | Trade Name: | EBV EA-D IgG ELISA Kit | |----------------------|----------------------------------| | Common Name: | EBV EA-D IgG EIA Test | | Classification Name: | EBV EA-D IgG Serological Reagent | 60-(0)7-3357-1222 ## Equivalent Device: Trinity Biotech Captia™ EBV EA-D IgG ELISA Device Description: The EBV EA-D IgG ELISA Kit is an enzyme-linked immunosorbent assay (ELISA) for the detection of IgG antibodies to EBV EA-D antigen in human serum. Intended Use: The Epstein Barr Virus Early Diffuse Antigen (EBV EA-D) IgG ELISA Test is for the qualitative detection of IgG antibodies to EBV EA-D as an aid in the diagnosis of EBV infection in patients with clinical symptoms of Infectious Mononucleosis (IM). The PANBIO EBV EA-D IgG ELISA should be used in conjunction with other EBV serologies. ## Principle of Procedure: Serum antibodies of the IgG class, when present, combine with EBV Early Antigen, which is purified by immunoaffinity chromatography, and attached to the polystyrene surface of the microwells. Residual serum is removed by washing and peroxidase conjugated anti-human IgG is added. The microwells are washed and a colourless substrate system, tetramethylbenzidine/ hydrogen peroxide (TMB/H2/22) 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. Colour development is indicative of the presence of EBV EA-D IgG antibodies in the test sample. {1}------------------------------------------------ ## PERFORMANCE CHARACTERISTICS ## Study Site 2: 346 frozen retrospective sera of various ages and genders were submitted to a state health laboratory in Maryland USA for EBV testing. The sera include samples from the following groups: 52 seronegative samples, 51 samples from patients with acute Infectious Mononucleosis, and 243 samples from patients with past exposure to EBV. These sera were tested on the PANBIO EBV EA-D IgG ELISA and the Trinity Biotech EBV EA-D IgG ELISA. The PANBIO 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 (table 1). Additionally, the Trinity Biotech results were compared to the EBV serological status (table 2), and the PANBIO versus the Trinity Biotech results (table 3) and are summarised below. | PANBIO ELISA | | | | | |--------------------------------------------------------------|----------|-----------|----------|-------| | EBV Status | Positive | Equivocal | Negative | Total | | Seronegative<br>VCA IgG (-)<br>VCA IgM (-)<br>EBNA IgG (-) | 0 | 1 | 51 | 52 | | Acute<br>VCA IgM (+)<br>EBNA IgG (-) | 16 | 6 | 29 | 51 | | Past Infection<br>VCA IgG (+)<br>VCA IgM (-)<br>EBNA IgG (+) | 60 | 22 | 161 | 243 | | Total | 76 | 29 | 241 | 346 | ## TABLE 1 EBV STATUS VERSUS PANBIO ELISA ## 95% Confidence Interval | Relative Sensitivity (Acute) | = 16/51 = 31.4 % | 19.1 – 45.9 % | |---------------------------------|--------------------|----------------| | Relative Sensitivity (Past) | = 60/243 = 24.7% | 19.3 – 30.1% | | Relative Specificity (Past) | = 161/243 = 66.3 % | 60.3 – 72.2 % | | Relative Specificity (Negative) | = 51/52 = 98.1 % | 89.7 – 100.0 % | | Relative Agreement | = 228/346 = 65.9 % | 60.9 – 70.9 % | *Retesting of equivocal samples was not conducted, as the samples were unavailable. {2}------------------------------------------------ | PANBIO ELISA | | | | | |--------------------------------------------------------------|----------|-----------|----------|-------| | EBV Status | Positive | Equivocal | Negative | Total | | Seronegative<br>VCA IgG (-)<br>VCA IgM (-)<br>EBNA IgG (-) | 4 | 2 | 46 | 52 | | Acute<br>VCA IgM (+)<br>EBNA IgG (-) | 29 | 6 | 16 | 51 | | Past Infection<br>VCA IgG (+)<br>VCA IgM (-)<br>EBNA IgG (+) | 92 | 21 | 130 | 243 | | Total | 125 | 29 | 192 | 346 | ## TABLE 2 EBV STATUS VERSUS TRINITY BIOTECH ELISA **PANBIO ELISA** 95% Confidence Interval | Relative Sensitivity (Acute) | = 29/51 | = 56.9% | 42.2 – 70.7% | |---------------------------------|-----------|---------|--------------| | Relative Sensitivity (Past) | = 92/243 | = 37.9% | 31.8 – 44.0% | | Relative Specificity (Past) | = 130/243 | = 53.5% | 47.2 – 59.8% | | Relative Specificity (Negative) | = 46/52 | = 88.5% | 76.6 – 95.6% | | Relative Agreement | = 205/346 | = 59.2% | 54.1 – 64.4% | *Retesting of equivocal samples was not conducted, as the samples were unavailable. ## TABLE 3 TRINITY BIOTECH VERSUS PANBIO ELISA | PANBIO ELISA | | | | | | |----------------|----------|-----------|----------|-------|--| | Trinity Result | Positive | Equivocal | Negative | Total | | | Positive | 72 | 16 | 37 | 125 | | | Equivocal | 2 | 4 | 23 | 29 | | | Negative | 2 | 9 | 181 | 192 | | | Total | 76 | 29 | 241 | 346 | | ## 95% Confidence Interval | Relative Sensitivity | = 72/125 | = 57.6% | 48.9 – 66.3% | |----------------------|-----------|----------|--------------| | Relative Specificity | = 181/192 | = 94.3 % | 90.0 – 97.1% | | Relative Agreement | = 253/346 | = 73.1% | 68.5 – 77.8% | *Retesting of equivocal samples was not conducted, as the samples were unavailable. {3}------------------------------------------------ ## Study Site 3: 330 prospective sera of various ages and genders were tested at a private pathology laboratory in Queensland Australia for EBV testing. The sera include the following groups: 47 seronegative, 35 with acute infectious mononucleosis and 248 with past exposure to EBV. These sera were tested on the PANBIO EBV EA-D IgG ELISA and the Trinity Biotech EBV EA-D IgG ELISA. The PANBIO results were compared to the Sera to determine the sensitivity, specificity, and agreement of the assay relative to the EBV serological status (table 4). Additionally, the Trinity Biotech results were compared to the EBV serological status (table 5), and the PANBIO versus the Trinity Biotech results (table 6) and are summarised below. | PANBIO ELISA | | | | | |----------------------------------------------------------------------------|----------|-----------|----------|-------| | EBV Status | Positive | Equivocal | Negative | Total | | Seronegative<br>VCA IgG (-)<br>VCA IgM (-)<br>EBNA IgG (-) | 2 | 0 | 45 | 47 | | Acute<br>VCA IgM (+)<br>EBNA IgG (-) | 13 | 0 | 22 | 35 | | Past Infection<br>VCA IgG (+)<br>VCA IgM (-)<br>EBNA IgG (+) | 33 | 1* | 214 | 248 | | Total | 48 | 1 | 281 | 330 | | Relative Sensitivity (Acute) = 13/35 = 37.1% 95% Confidence In 21.5 - 55.1 | | | | | ## TABLE 4 EBV STATUS VERSUS PANBIO ELISA | | | | 95% Confidence Interval | |---------------------------------|-----------|---------|-------------------------| | Relative Sensitivity (Acute) | = 13/35 | = 37.1% | 21.5 - 55.1% | | Relative Sensitivity (Past) | = 33/248 | = 13.3% | 9.1 - 17.5% | | Relative Specificity (Past) | = 214/248 | = 86.3% | 82.0 - 90.6% | | Relative Specificity (Negative) | = 45/47 | = 95.7% | 85.5 - 99.5% | | Relative Agreement | = 272/330 | = 82.4% | 78.3 - 86.5% | *Equivocal samples were inconclusive following repeat testing on the alternative method (IFA). {4}------------------------------------------------ | TABLE 5 | | | | | |---------|--|--|-----------------------------------------|--| | | | | EBV STATUS VERSUS TRINITY BIOTECH ELISA | | | PANBIO ELISA | | | | | |--------------------------------------------------------------|----------|-----------|----------|-------| | EBV Status | Positive | Equivocal | Negative | Total | | Seronegative<br>VCA IgG (-)<br>VCA IgM (-)<br>EBNA IgG (-) | 1 | 0 | 46 | 47 | | Acute<br>VCA IgM (+)<br>EBNA IgG (-) | 21 | 0 | 14 | 35 | | Past Infection<br>VCA IgG (+)<br>VCA IgM (-)<br>EBNA IgG (+) | 38 | 1* | 209 | 248 | | Total | 60 | 1 | 269 | 330 | | | | | 95% Confidence Interval | |---------------------------------|-----------|---------|-------------------------| | Relative Sensitivity (Acute) | = 21/35 | = 60.0% | 42.1 – 76.1% | | Relative Sensitivity (Past) | = 38/248 | = 15.3% | 10.8 – 19.8% | | Relative Specificity (Past) | = 209/248 | = 84.3% | 79.7 – 88.8% | | Relative Specificity (Negative) | = 46/47 | = 97.9% | 88.7 – 99.9% | | Relative Agreement | = 276/330 | = 83.7% | 79.8 – 87.7% | *Equivocal samples were inconclusive following repeat testing on the alternative method (IFA). | TABLE 6 | |-------------------------------------| | TRINITY BIOTECH VERSUS PANBIO ELISA | | PANBIO ELISA | | PANBIO ELISA | | | | | |----------------------|----------|-----------|----------|-------------------------| | Trinity Biotech | Positive | Equivocal | Negative | Total | | Positive | 35 | 1 | 24 | 60 | | Equivocal | 0 | 0 | 1 | 1 | | Negative | 13 | 0 | 256 | 269 | | Total | 48 | 1 | 281 | 330 | | | | | | 95% Confidence Interval | | Relative Sensitivity | | = 35/60 | = 58.3% | 44.9 - 70.9% | | Relative Specificity | | = 256/269 | = 95.2% | 91.9 - 97.4% | | Relative Agreement | | = 291/330 | = 88.2% | 84.7 - 91.7% | *Equivocal samples were inconclusive following repeat testing on the alternative method (IFA). {5}------------------------------------------------ ## REPRODUCIBILITY ## Study Sites 3, 4 & 5: The reproducibility of the PANBIO EBV EA-D IgG 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 site and total precision were estimated by analysis of variance (ANOVA Type II). The results are presented in table 7 below. ## TABLE 7 -- Reproducibility Data PANBIO EBV EA-D IgG Study Sites 1, 2 & 3 | Sample | n | *Mean | Within | | Between Day | | Between Site | | Total | | |----------|----|-------|--------|-------|-------------|------|--------------|------|-------|-------| | | | | *S.D | CV | *S.D | CV | *S.D | CV | *S.D | CV | | Positive | 27 | 1.94 | 0.39 | 20.3% | 0.00 | 0.0% | 0.08 | 4.3% | 0.39 | 20.1% | | Cut-off | 27 | 1.00 | 0.11 | 10.9% | 0.00 | 0.0% | 0.00 | 0.0% | 0.10 | 10.1% | | Negative | 27 | 0.11 | 0.02 | 18.9% | 0.00 | 0.0% | 0.01 | 5.8% | 0.02 | 19.4% | | 1 | 27 | 3.78 | 0.34 | 9.0% | 0.00 | 0.0% | 0.16 | 4.1% | 0.36 | 9.4% | | 2 | 27 | 0.94 | 0.11 | 11.7% | 0.02 | 2.4% | 0.04 | 4.7% | 0.12 | 12.5% | | 3 | 27 | 1.61 | 0.17 | 10.8% | 0.00 | 0.0% | 0.00 | 0.0% | 0.17 | 10.4% | | 4 | 27 | 0.87 | 0.09 | 10.9% | 0.00 | 0.0% | 0.00 | 0.0% | 0.09 | 10.2% | | 5 | 27 | 1.26 | 0.18 | 14.0% | 0.00 | 0.0% | 0.04 | 3.3% | 0.17 | 13.9% | | 6 | 27 | 2.51 | 0.30 | 12.2% | 0.07 | 2.7% | 0.00 | 0.0% | 0.31 | 12.2% | | 7 | 27 | 1.16 | 0.11 | 9.5% | 0.11 | 9.1% | 0.06 | 5.5% | 0.15 | 13.0% | | 8 | 27 | 1.12 | 0.16 | 14.0% | 0.11 | 9.9% | 0.04 | 3.7% | 0.19 | 16.5% | #### Precision Measures (Using Ratio) SD = Standard Deviation CV = Coefficient of Variation (%) * Values calculated from ratios Site 1: Three days of triplicates Site 2: Three days of triplicates Site 3: Three days of triplicates ## Note: Standard Deviation results have been rounded to two decimal places for tabulation purposes. {6}------------------------------------------------ ## POTENTIAL CROSS-REACTIVITY ## Study Site 5: This study consisted of a panel of 50 specimens screened for IgG antibodies detectable by ELISA to disease types other than Epstein Barr Virus. The purpose of this study is to establish the analytical specificity of the EBV EA-D IgG ELISA Test, through the analysis of specimens 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 of the specimens with the EBV EA-D IgG ELISA Test. Table 9 on the following page lists the cross-reactivity results for each type of specimen included in the disease panel. Table 8 below provides a summary of the cross-reactivity data. ## TABLE 8 - PANBIO EBV EA-D IgG | Disease (IgG Antibodies) | Total Specimens | Positive Result | |--------------------------|-----------------|-----------------| | Cytomegalovirus | 10 | (0/10) | | Varicella zoster | 13 | (0/13) | | Herpes simplex virus 1 | 8 | (0/8) | | Herpes simplex virus 2 | 2 | (0/2) | | Anti-Nuclear Antibody | 8 | (0/8) | | Rheumatoid Factor | 9 | (0/9) | | Total Antibody | 50 | (0/50) | ## CROSS-REACTIVITY SPECIMEN PANEL SUMMAY Results indicate that no specimens (0/50) were positive when analysed with the EBV EA-D IgG ELISA Kit. The true negative result of 100% for the above disease panel is consistent with good analytical specificity for the EBV EA-D IgG ELISA Test. {7}------------------------------------------------ #### DEPARTMENT OF HEALTH & HUMAN SERVICES Image /page/7/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a stylized eagle with three wing-like shapes extending from its head. The eagle faces right and is positioned within a circular border containing the text "DEPARTMENT OF HEALTH & HUMAN SERVICES USA". Food and Drug Administration 098 Gaither Road Rockville MD 20850 SEP 27 2002 Ms. Kate Wersin Regulatory Affairs Officer PANBIO Limited 116 Lutwyche Road Windsor Brisbane, Queensland Australia 4030 Re: k021793 Trade/Device Name: EBV-EA-D IgG ELISA Regulation Number: 21 CFR 866.3235 Regulation Name: Epstein-Barr Virus Serological Reagents Regulatory Class: Class I Product Code: LSE Dated: August 12, 2002 Received: August 15, 2002 Dear Ms. Wersin: 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, 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 (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, {8}------------------------------------------------ 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 (301) 594-4588. Additionally, for questions on the promotion and advertising of your device, please contact the Office of Compliance at (301) 594-4639. 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 (800) 638-2041 or (301) 443-6597 or at its internet address "http://www.fda.gov/cdrh/dsma/dsmamain.html". Sincerely yours, Steven Butman 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 {9}------------------------------------------------ Page 1 of 1 510(k) Number (if known): K021793 Device Name: Indications For Use: The Epstein Barr Virus Early Diffuse Antigen (EBV EA-D) IgG ELISA Test is for the qualitative detection of IgG antibodies to EBV EA-D as an aid in the diagnosis of EBV infection in patients with clinical symptoms of Infectious Mononucleosis (IM). The PANBIO EBV EA-D IgG ELISA should be used in conjunction with other EBV serologies. (PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED) Concurrence of CDRH, Office of Device Evaluation (ODE) Worley Dubose --- (Division Sign Off) sion Sign-Laboratory Dev Division of Clink 510(k) Number (Optional Format 3-10-98)