EA-D IGM ELISA TEST SYSTEM

{0}------------------------------------------------ # NOV 2 5 1998 K982352 # 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: | March 23, 1998 | |----------------|------------------------------------------------------| | Name: | Columbia Bioscience, Inc. | | Address: | 8775 M Centre Park Drive, #559<br>Columbia, MD 21045 | | Contact Person: | Norman Jenkins | |-----------------|----------------| | PhoneNumber. | 410-995-0450 | | Fax Number. | 410-995-0448 | #### Device Information: | Trade Name: | EA-D IgM ELISA Kit | |----------------------|--------------------| | Common Name. | EA-D IgM EIA Test | | Classification Name; | Epstein-Barr Virus | Equivalent Device: EBV Serology Device Description: The & EA-D IgM ELISA Kit is an enzyme-linked immunosorbent assay (ELISA) for the detection of IgM antibodies to Epstein-Barr Early antigen diffuse in human serum. Intended Use: For the qualitative determination of IgM antibodies in human serum to Epsien Barr Virus (recombinant) Early antigen (EA-D) antigen. The & EA-D IgG assay should be used in conjunction with other Epstein-Barr serologies (VCA IgM. VCA IgG. EBNA-1 IgM and heterophile) as an aid in the diagnosis of infectious mononucleosis. The test can be performed either manually or in conjunction with the MAGO PLUS TM Automated EIA Processor. Principle of Procedure: The & EA-D IgG ELISA Kit is an enzyme-linked immunosorbent assay to detect IgG to EA-D in human serum. Recombinant EA-D antigen is attached to a solid phase (microtiter well). Diluted test sera are added to each well. If antibodies which recognize the. EA-D antigen are present in the patient sample they will bind to the antigen in the well. After incubation, the wells are washed to remove unbound antibody. An enzyme labeled anti-human immunoglobulin (conjugate) is added to each test well. If antibody is present the enzyme-linked antibody will bind to it. After incubation, the wells are washed to remove unbound conjugate. A substrate solution is then added to each well. If enzyme is present from prior step, the reaction is stopped and the, color intensity is measured photometrically producing an indirect detection of the specific antibody present in the patient sample. {1}------------------------------------------------ # Performance Characteristics # A. Clinical Sensitivity and Specificity Using Characterized Sera Frozen retrospective sera from one hundred and seventy-six patients were characterized using commercially available kits for VCA IgM, VCA IgG, EBNA IgG and heterophile antibodies. Based on the results of this testing, the patient sera were characterized as follows: * 102 sera were characterized as convalescent (past infection). These were positive for VCA IgG and/or EBNA IgG antibodies and negative for VCA IgM and heterophile antibody. * 32 sera were characterized as scronegative. These were negative for VCA IgG, VCA IgM, EBNA IgG and heterophile antibody. * 42 sera were characterized as having a current (recent) infection. These were positive for VCA IgM and/or heterophile antibody and were negative for EBNA IgG. All 176 sera were then tested by an independent clinical commercial laboratory using the Is-EBV-EA-D IgM Test Kit. The results obtained are shown in Table 2: | | TABLE 2 | <i>EBV Serological Status</i> | | | |-----------------|------------|-------------------------------|-------------------|--------------| | | | Convalescent | Current Infection | Seronegative | | | POSITIVE | 5 | 17 | . 1 | | Is-EBV-EA-D-IgM | NEGATIVE | 93 | 20 | 29 | | | *EQUIVOCAL | 4 | 5 | 2 | | | | | 95% CI | |------------------------------------------|---------|---------|------------| | Relative Specificity (Convalescent) | 93/98 | = 94.9% | 88.5-98.3% | | Relative Sensitivity (Current Infection) | 17/37 | = 45.9% | 29.5-63.1% | | Relative Specificity (Seronegative) | 29/30 | = 96.7% | 82.8-99.9% | | Overall Agreement | 139/165 | = 84.2% | 78.7-89.8% | # ERV Serological Status * Equivocal results were excluded from calculations NOTE : Please be advised that 'relative' refers to the comparison of the 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's accuracy to predict disease. Since the above studies were performed on a pre-selected, retrospective population, no calculations for the assay's positive predictive value may be done or inferred. {2}------------------------------------------------ # B.Precision Todclerminethe precisionofthels-EBV-EA-DIgMTestKit, fourpositive and wonegativesera were assayed ten timescachinthreedifferentrunsatthreedifferentsites. The3sitesinclude: themanufacturer, arescarchanddevelopmentlaboratory,andaclinicalcommerciallaboratory. Theintra-andinterassayprecisionobtainedateachsiteis showninTables3,4and5.TheInter-SiteprecisionisshowninTable6. | SERUM | INTRA-ASSAY RUN 1 | | INTRA-ASSAY RUN 2 | | INTRA-ASSAY RUN 3 | | INTERASSAY | | | |---------|-------------------|-------|-------------------|-------|-------------------|-------|------------|-------|-------| | | MEAN INDEX | CV% | MEAN INDEX | CV% | MEAN INDEX | CV% | MEAN INDEX | CV% | | | A (POS) | 6.59 | 5.42 | 6.37 | 4.53 | 6.25 | 5.17 | 6.40 | 5.35 | | | B (POS) | 1.86 | 12.60 | 1.79 | 10.84 | 1.95 | 6.18 | 1.87 | 10.46 | | | C (POS) | 2.90 | 10.05 | 2.67 | 7.31 | 2.61 | 9.00 | 2.73 | 9.77 | | | D (POS) | 1.37 | 8.92 | 1.38 | 12.45 | 1.41 | 12.81 | 1.39 | 11.20 | | | E (NEG) | 0.14 | 44.84 | 0.15 | 39.58 | 0.16 | 46.08 | 0.15 | 42.74 | | | F (NEG) | 0.44 | 15.23 | 0.45 | 39.42 | 0.32 | 35.23 | 0.40 | 34.16 | | | | | | | | | CAL | 0.97 | 10.49 | n = 9 | | | | | | | | PC | 1.65 | 12.75 | n = 3 | | | | | | | | NC | 0.29 | 38.40 | n = 3 | TABLE 4 : Site #2- Intra-Assay and Interassay Precision | SERUM | | INTRA-ASSAY RUN 1 | INTRA-ASSAY RUN 2 | INTRA-ASSAY RUN 3 | INTERASSAY | | | | | |---------|---------------|-------------------|-------------------|-------------------|------------|---------------|-------|---------------|-----| | | MEAN<br>INDEX | MEAN<br>INDEX | CV% | MEAN<br>INDEX | CV% | MEAN<br>INDEX | CV% | MEAN<br>INDEX | CV% | | A (POS) | 6.328 | 6.78 | 5.514 | 4.34 | 6.055 | 3.85 | 5.966 | 7.68 | | | B (POS) | 1.813 | 9.32 | 1.580 | 10.07 | 1.722 | 6.65 | 1.705 | 10.21 | | | C (POS) | 2.465 | 5.70 | 2.376 | 17.21 | 2.521 | 6.15 | 2.454 | 10.72 | | | D (POS) | 1.261 | 4.89 | 1.118 | 6.07 | 1.219 | 3.78 | 1.199 | 6.97 | | | E (NEG) | 0.144 | 28.01 | 0.155 | 16.49 | 0.150 | 14.21 | 0.149 | 19.71 | | | F (NEG) | 0.289 | 15.47 | 0.341 | 8.87 | 0.308 | 10.00 | 0.313 | 13.11 | | | | | | | | CAL | 1.001 | 4.69 | n = 18 | | | | | | | | PC | 1.564 | 12.03 | n = 12 | | | | | | | | NC | 0.303 | 4.72 | n = 12 | | | | | | TABLE 5 : Site #3 - Intra-assay and Interassay Precision | | |--|--|--|----------------------------------------------------------|--| |--|--|--|----------------------------------------------------------|--| | SERUM | INTRA-ASSAY RUN 1 | | INTRA-ASSAY RUN 2 | | INTRA-ASSAY RUN 3 | | INTERASSAY | | |---------|-------------------|-------|-------------------|-------|-------------------|-------|---------------|-------| | | MEAN<br>INDEX | CV% | MEAN<br>INDEX | CV% | MEAN<br>INDEX | CV% | MEAN<br>INDEX | CV% | | A (POS) | 6.35 | 11.01 | 6.53 | 12.63 | 6.05 | 11.29 | 6.31 | 11.74 | | B (POS) | 1.98 | 9.08 | 1.90 | 10.74 | 1.91 | 11.87 | 1.93 | 10.36 | | C (POS) | 2.63 | 16.02 | 2.59 | 15.59 | 2.52 | 17.40 | 2.58 | 15.86 | | D (POS) | 1.55 | 20.22 | 1.49 | 26.10 | 1.42 | 14.80 | 1.49 | 20.67 | | E (NEG) | 0.13 | 15.27 | 0.12 | 19.99 | 0.13 | 23.17 | 0.13 | 19.85 | | F (NEG) | 0.29 | 15.16 | 0.24 | 15.18 | 0.36 | 10.88 | 0.29 | 21.96 | | | | | | | | CAL | 1.00 | 11.59 | | | | | | | | PC | 1.42 | 6.45 | | | | | | | | NC | 0.30 | 5.77 | #### Table 6 : Inter-Site Precision | SERUM<br>(n=90) | INTER-SITE | | |-----------------|---------------|-------| | | MEAN<br>INDEX | CV% | | A (POS) | 6.23 | 9.10 | | B (POS) | 1.83 | 11.51 | | C (POS) | 2.58 | 12.97 | | D (POS) | 1.36 | 17.32 | | E (NEG) | 0.14 | 31.41 | | F (NEG) | 0.34 | 30.28 | | CAL (n=36) | 0.99 | 8.21 | | LPC (n=18) | 1.55 | 11.87 | | NC (n=18) | 0.30 | 13.42 | {3}------------------------------------------------ ### C. Specificity with Potentially Cross-Reactive Sera Thirteen sera, non-reactive (negative) for IgM antibodies to EA-D in the Is-EBV-EA-D IgM Test Kit, were tested by EIA for IgM antibody to varicclla zoster, cytomegalovirus and herpes simplex virus. 4/4 anti-VZV IgM positive scra were non-reactive for anti-EA-D IgM; 3/5 anti-CMV IgG positive sera were non-reactive for anti-EA-D IgM and 4/4 anti-HSV positive scra were non-reactive for anti-EA-G IgM. This suggests that some cross-reactivity should be expected with the Is-FA-D IgM Test Kit from these analytes. #### D. Correlation of Manual and MAGO Plus Results The Is-I'D V-EA-D IgM Test Kit has been developed for automated as well as manual use. To demonstrate the equivalence of the manual and MAGO Plus procedures, the results of 128 serum samples tested by both methods were plotted. A scattergram and regression line of the results obtained with 95% confidence intervals is shown in Figure 3. The data indicate good correlation with a Pearson Correlation Coefficient of 0.975. Image /page/3/Figure/4 description: This image is a scatter plot comparing "MAGO PLUS INDEX VALUES" on the y-axis and "MANUAL INDEX VALUES" on the x-axis. The data points are clustered around a regression line, indicating a positive correlation between the two variables. The equation of the regression line is given as Y = 0.0657 + 0.9423 X, and the correlation coefficient (r) is 0.975, suggesting a strong linear relationship. FIGURE 3 : Manual and MAGO Plus Result Correlation ### E. MAGO Plus Precision The precision of the assay when performed on the MAGO Plus Automated EIA Processor was determined by assaying six sera ten times each in three different runs. Table 7 shows the intra-and interassay precision obtained using the MAGO Plus. TABLE 7 : Site #2- Intra-Assay and Interassay Precision - MAGO Plus | SERUM | INTRA-ASSAY RUN 1<br>MEAN<br>INDEX | INTRA-ASSAY RUN 1<br>CV% | INTRA-ASSAY RUN 2<br>MEAN<br>INDEX | INTRA-ASSAY RUN 2<br>CV% | INTRA-ASSAY RUN 3<br>MEAN<br>INDEX | INTRA-ASSAY RUN 3<br>CV% | INTERASSAY<br>MEAN<br>INDEX | INTERASSAY<br>CV% | |---------|------------------------------------|--------------------------|------------------------------------|--------------------------|------------------------------------|--------------------------|-----------------------------|-------------------| | A (POS) | 6.1 | 3.76 | 5.8 | 4.29 | 5.8 | 3.98 | 5.9 | 4.35 | | B (POS) | 1.8 | 6.38 | 1.7 | 7.27 | 1.8 | 8.12 | 1.8 | 7.44 | | C (POS) | 2.7 | 5.51 | 2.5 | 7.23 | 2.6 | 7.49 | 2.6 | 6.97 | | D (POS) | 1.4 | 16.10 | 1.3 | 7.77 | 1.3 | 8.34 | 1.3 | 11.76 | | E (NEG) | 0.1 | 37.16 | 0.1 | 28.75 | 0.1 | 0.00 | 0.1 | 30.51 | | F (NEG) | 0.3 | 23.80 | 0.3 | 20.45 | 0.3 | 15.06 | 0.3 | 20.86 | | | | | | | | CAL | 1.0 | 2.80 | | | | | | | | PC | 1.7 | 3.50 | | | | | | | | NC | 0.4 | 0.00 | {4}------------------------------------------------ Image /page/4/Picture/1 description: The image shows the seal of the Department of Health & Human Services (HHS) of the United States of America. The seal features the department's name encircling a symbol. The symbol consists of a stylized caduceus, which is a traditional symbol of medicine, with three figures representing health and human services. The text is arranged in a circular pattern around the symbol. Food and Drug Administration 2098 Gaither Road Rockville MD 20850 NOV 25 1998 Norman Jenkins President Columbia Bioscience, Inc. 8775 M Centre Park Drive, #559 Columbia, Maryland 21045 Re: K982350 Device: EA-D IgM ELISA Test System Dated: September 22, 1998 Received: September 28, 1998 Dear Mr. Jenkins: 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 OS 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. {5}------------------------------------------------ #### 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 (770)488-7655. 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 (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" (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 (800) 638-2041 or at (301) 443-6597 or at its internet address "http://www.fda.gov/cdrh/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 {6}------------------------------------------------ Page 1 of 1 510(k) Number: Not Known Device Name: & EA-D IgM ELISA Indications For Use: For the qualitative determination of IgM antibodies in human serum to Epstein Barr (recombinant) early antigen diffuse (EA-D) antigen. The & EA-D IgM assay should be used in conjunction with other Epstein-Barr serologies (VCA IgM, VCA IgG, EBNA-1 IgG, EBNA-1 IgM, EA-D IgG and heterophile) as an aid in the diagnosis of infectious mononucleosis. The test can be performed either manually or in conjunction with the MAGO PLUS The Automated EIA Processor. #### PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED) ------------------------------------------------------------------------------------------------------------------------------------------------------------------------------ Concurrence of CDRH, Office of Device Evaluation (ODE) Prescription Use (Per 21 CFR 801.109) OR Over-The Counter Use (Optional Format 1-2-96) Woody Dubois inical I aboratory Devices 510(k) Number