DIAMEDIX IS-ANTI-CARDIOLIPIN SCREEN TEST SYSTEM

{0}------------------------------------------------ # AUG 2 0 2001 K012053 ## 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: ## Applicant Information: | Date Prepared: | June 29, 2001 | |----------------|----------------------| | Name: | Diamedix Corporation | | Address: | 2140 N. Miami Avenue | | | Miami, FL 33127 | | Contact Person: | Dr. Lynne Stirling | |-----------------|--------------------| | Phone Number: | 305-324-2354 | | Fax Number: | 305-324-2388 | #### Device Information: | Trade Name: | Is anti-Cardiolipin Screen Test System | |----------------------|-------------------------------------------| | Common Name: | Anti-Cardiolipin ELISA test | | Classification Name: | Anticardiolipin immunological test system | #### Equivalent Device: Orgentec Anti-cardiolipin Screen ELISA Assay Device Description: The Is anti-Cardiolipin Screen Test System is an enzyme-linked immunosorbent assay (ELISA) for the semi-quantitative measurement of IgG, IgM and IgA antibodies to cardiolipin in human serum Intended Use: The assay is intended for the semi-quantitative measurement of IgG. IgM and IgA antibodies to cardiolipin in human serum. The results of the assay can be used as an aid in the assessment of the risk of thrombosis in patients with SLE or SLE-like dosorders. #### Principle of the Procedure: The Is-anti-Cardiolipin Screen Test System is an indirect solid-phase enzyme immunoassay. Highly purified cardiolipin is coated onto plastic microwells and saturated with highly purified human B2-Glycoprotein I. Controls and diluted patient samples are added to the wells. Any patient IgG, IgM or IgA antibodies in the sample bind to the well. Anti-human horseradish peroxidase conjugate is then added After incubation and washing, a substrate solution is then added to each well. In the presence of bound enzyme, the substrate is converted to a blue colored product. After acid additon to stop the reaction, a yellow end product is formed that is read spectrophotometrically at 450 nm (reference 600-630 mm) and is directly proportional to the concentration of cardiolipin IgG, IgM and IgA antibodies in the sample. {1}------------------------------------------------ # SUMMARY OF SAFETY AND EFFECTIVENESS ## Performance Characteristics ## A. Relative Sensitivity and Specificity Two hundred and three frozen, retrospective sera were tested for IgG/IgM/IgA cardiolipin antibodies using the I wo hand careen Test Kit and a commercially available ELISA kit for detecting cardiolipin IgG/IgM/IgA antibodies. Based on the results of this resing the relative sensitivity, relative specificity and overall agreement were calculated. The results obtained are shown below. Further resolution of the discordant samplesshowed that were samples that were negative in the Is anti-Cardiolipin Screen and positive by the other EIA were negative by a referee EIA method. The remaining ten discordant samples were positive in the referee test. Is-anti-Cardiolipin Screen | | | Positive | Negative | *Equivocal | |------------------------------------------------------------------|------------|----------|----------|-------------| | Other<br>EIA | Positive | 83 | 18 | 3 | | | Negative | 0 | 99 | 0 | | | *Equivocal | 0 | 0 | 0 | | **95% CI | | | | | | Relative Sensitivity | | 83/101 | = 82.2 % | 74.7-89.6% | | Relative Specificity | | 99/99 | = 100.0% | 96.3-100.0% | | Overall Agreement | | 182/200 | = 91.0% | 86.2-94.6% | | * Equivocal results were excluded from calculations. | | | | | | ** 95% Confidence Intervals (CI) calculated by the Exact Method. | | | | | NOTE : Please be advised that '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 judgement can be made on the comparison's accuracy to predict disease. ## B. Clinical Sensitivity and Specificity Diseases A total of three hundred and forty-five frozen retrospective, clinically characterized sera were assayed using the Is anti-Cardiolipin Screen Test Kit in order to assess both the clinical sensitivity and clinical specificity of the test system. These samples consisted of 215 normal sera, 57 sera from patients with diagnosed anti-phospholipid syndrome (APS), 34 sera from patients with systemic lupus erythematosus (SLE), 24 sera from patients with other autoimmune diseases such as Sjogren's Syndrome, scleroderma, polymyositis and rheumatoid arthritis and 15 samples from patients with positive RPR titers. Results are summarized below. Note that the all positive samples were also positive when tested by another commercially available ELISA test. | Patient Group | Total | Positive | Negative | Equivocal | |------------------------------|---------------|-----------|----------|-----------| | Normals | 215 | 9 | 205 | 1 | | APS | 57 | 54 | 3 | 0 | | SLE | 34 | 10 | 19 | 5 | | Other Autoimmune<br>Diseases | 24 | 6 | 18 | 0 | | RPR Positive | 15 | 5 | 10 | 0 | | | | | | | | Clinical Specificity: | # Neg/Total # | | | | | Normals | 205/215 | $=$ 95.3% | | | | RPR Positive | 10/15 | $=$ 66.7% | | | | Clinical Sensitivity : | # Pos/Total # | | | | | APS | 54/57 | $=$ 94.7% | | | | SLE | 10/34 | $=$ 29.4% | | | | Other Autoimmune | 6/24 | $=$ 25.0% | | | 000170 {2}------------------------------------------------ ## C. Correlation of Manual and MAGO Plus results The Is anti-Cardiolipin Screen 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 152 serum samples tested for anti-Cardiolipin IgG/IgM/IgA antibodies by both the manual and automated methods, and whose values were within the reportable range of the assay, were plotted. Scattergrams and regression lines of the results obtained with 95% confidence intervals are shown in FIGURE 1. The data indicate good correlation with a Correlation Coefficient (r) of 0.9497. Image /page/2/Figure/2 description: The image is a scatter plot titled "FIGURE 3: Is anti-Cardiolipin Screen Manual vs MAGO Plus Correlation". The x-axis is labeled "MANUAL U/ML" and ranges from 0 to 100. The y-axis is labeled "MAGO Plus U/ML" and ranges from 0 to 120. The plot shows a positive correlation between the two variables, with data points scattered around a regression line. ## D. Precision To assess the precision of the Is anti-Cardiolipin Screen Test Kit six serum samples of varying reactivity (two negative and four positive) were tested in three separate runs. Precision was assessed both manually and using the MAGO Plus Automated EIA Processor. The results obtained are shown in below. | SERUM | INTRA-ASSAY DAY 1 | | | INTRA-ASSAY DAY 2 | | | INTRA-ASSAY DAY 3 | | | INTERASSAY (n=9) | | | |-------|-------------------|-------|-------|-------------------|-------|-------|-------------------|-------|------|------------------|-------|-------| | | MEAN U/ml | SD | CV% | MEAN U/ml | SD | CV% | MEAN U/ml | SD | CV% | MEAN U/ml | SD | CV% | | A | 3.3 | 0.265 | 8.02 | 3.3 | 0.173 | 5.25 | 3.7 | 0.115 | 3.09 | 3.4 | 0.274 | 7.97 | | B | 2.8 | 0.404 | 14.61 | 3.3 | 0.058 | 1.77 | 3.8 | 0.252 | 6.57 | 3.3 | 0.521 | 15.83 | | C | 21.4 | 0.945 | 4.41 | 21.9 | 0.950 | 4.33 | 24.6 | 1.443 | 5.86 | 22.7 | 1.787 | 7.88 | | D | 33.4 | 3.317 | 9.94 | 36.0 | 0.700 | 1.94 | 34.7 | 1.411 | 4.07 | 34.7 | 2.161 | 6.23 | | E | 52.9 | 2.957 | 5.59 | 58.8 | 6.851 | 11.65 | 72.5 | 2.207 | 3.04 | 61.4 | 9.527 | 15.51 | | F | 40.1 | 1.528 | 3.81 | 44.8 | 0.954 | 5.83 | 45.0 | 1.473 | 3.27 | 43.3 | 2.654 | 6.13 | Manual Intra-Assay and Interassay Precision for Is-anti-Cardiolipin Screen MAGO Plus Intra-Assay and Interassay Precision for Is-anti-Cardiolipin Screen | SERUM | INTRA-ASSAY DAY 1 | | | INTRA-ASSAY DAY 2 | | | INTRA-ASSAY DAY 3 | | | INTERASSAY (n=9) | | | |-------|-------------------|-------|-------|-------------------|--------|-------|-------------------|--------|-------|------------------|--------|-------| | | MEAN<br>U/ml | SD | CV% | MEAN<br>U/ml | SD | CV% | MEAN<br>U/ml | SD | CV% | MEAN<br>U/ml | SD | CV% | | A | 4.6 | 0.058 | 1.25 | 6.8 | 0.503 | 7.44 | 9.8 | 1.762 | 17.91 | 7.1 | 2.442 | 34.51 | | B | 4.2 | 0.289 | 6.82 | 6.5 | 0.300 | 4.62 | 8.7 | 0.306 | 3.50 | 6.5 | 1.966 | 30.29 | | C | 27.0 | 1.115 | 4.13 | 36.6 | 2.600 | 7.10 | 36.2 | 5.101 | 14.08 | 33.3 | 5.555 | 16.70 | | D | 42.5 | 1.686 | 3.97 | 63.4 | 3.509 | 5.53 | 53.2 | 6.934 | 13.03 | 53.0 | 9.899 | 18.67 | | E | 79.3 | 7.927 | 10.00 | 92.2 | 19.931 | 21.62 | 74.9 | 11.243 | 15.00 | 82.1 | 14.390 | 17.52 | | F | 46.6 | 3.287 | 7.06 | 66.4 | 5.179 | 7.80 | 63.0 | 6.933 | 11.01 | 58.7 | 10.290 | 17.54 | Image /page/2/Picture/9 description: The image shows the number 000171 in a bold, sans-serif font. The numbers are printed in black ink on a white background. The numbers are slightly distorted, with some of the edges appearing blurred. The number is likely a serial number or some other type of identification number. {3}------------------------------------------------ # Expected Values The prevalence of anti-cardiolipin antibodies may vary depending on a number of factors such as age, gender, geographical location, race, type of test used and clinical history of individual patients. Antibodies to anti-cardiolipin are generally absent, or have a very low incidence, in the normal healthy population. Increased incidence can occur in the elderly population. A published study has shown a prevalence of 12% in the elderly population (mean age of 70 years) as opposed to 2% for a younger population. In addition, anti-cardiolipin antibodies were detected in 23% of elderly individuals who were also positive for anti-nuclear antibodies (12). In the present study, the expected values for a normal, healthy population were assessed by testing sera from one hundred and forty-eight S. Florida blood donors (ninety-eight males and fifty females) in the Is-anti-Cardiolipin Screen Test Kit. One hundred and forty sera (94.6%) were negative for antibodies, eight sera (5.4%) were positive and none were equivocal. The age distribution and antibody prevalence for this population are shown in the table below. The expected values for a clinical population were assessed by testing fifty-seven sera from patients with a diagnosis of anti-phospholipid syndrome (APS) in the Is-anti-Cardiolipin Screen Test Kit. Fifty-four (94.7%) were positive, three (5.3%) were negative and none were equivocal for IgG/IgM/ IgA antibodies. Histograms showing the distribution of values for these normal and clinical populations are shown in FIGURES 1 and 2. | | Number of Donors | Prevalence | |-------------------------|---------------------|------------| | Total Number | 148 | | | Geographic<br>Location: | South Florida : 148 | 5.4% | | Age | | | | 10-19 | 7 | 14.3% | | 20-29 | 36 | 0.0% | | 30-39 | 73 | 8.2% | | 40-49 | 22 | 4.5% | | 50-59 | 8 | 0.0% | | 60-69 | 2 | 0.0% | #### Age Distribution and Prevalence of anti-Cardiolipin IgG/IgM/IgA in a Normal S. Florida Population {4}------------------------------------------------ FIGURE 1 Distribution of anti-Cardiolipin IgG/IgM/IgA in a Normal Population Image /page/4/Figure/1 description: This image is a bar graph that shows frequency on the y-axis and U/ML on the x-axis. The frequency ranges from 0 to 70, and the U/ML ranges from 0 to 60. There are two bars that are significantly higher than the rest, one at 0 U/ML with a frequency of 70, and one at 5 U/ML with a frequency of 64. FIGURE 2 Distribution of anti-CardiolipinlgG/lgM/ IgA in a Clinical Population Image /page/4/Figure/3 description: This image is a histogram showing frequency on the y-axis and U/ML on the x-axis. The frequency ranges from 0 to 7, and the U/ML ranges from 0 to 280. The histogram shows the distribution of U/ML values, with the highest frequencies occurring around 80 and 160. {5}------------------------------------------------ DEPARTMENT OF HEALTH & HUMAN SERVICES 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 circle with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. Inside the circle is a stylized image of an eagle with three heads. AUG 2 0 2001 Food and Drug Administration 2098 Gaither Road Rockville MD 20850 Lynne Stirling, Ph. D. Diamedix Corporation 2140 N. Miami Avenue Miami, Florida 33127 Re: K012053 K012055 Trade/Device Name: Diamedix Is-anti-Cardiolipin Screen Test System Regulation Number: 21 CFR 866.5660 Regulatory Class: Class II Product Code: MID Dated: June 29, 2001 Received: July 2, 2001 Dear Dr. Stirling: We have reviewed your Section 510(k) notification of intent to market the device referenced in we have leviewed your becaon b ro(x) x = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = = above and we nave determinou the act 1976, the enactment date of the Medical Device intersiale comments, or to devices that have been reclassified in accordance with the provisions of the same issues and issues and issues and issues and issues and issues and Amendments, or to devices mat have been rockes market the device, subject to Federal Food, Drug, and Cosmons Free (10) - 100 general controls provisions of the Act include the general controls provisions of the Freis - Fittigential of the gractice, labeling, and prohibitions against misbranding and adulteration. If your device is classified (see above) into either class II (Special Controls) or class III If your device is classifica (see above) into such additional controls. Existing major regulations (Premarket Approval), it may be subject to ade a subject to address, Title 21, Parts 800 to 895. affecting your device can be found in the Code of Federal Regulations, Title A substantially equivalent determination assumes compliance with the Good Manufactures a A substantally equivalent determination as and and and the mart 820) and that, through Fractice for Mculear Dovies: Seneral (Concertation (FDA) will verify such periodic GMT inspections, the Food and Suns and assumptions. Tantife to comply with the concerning your device in the Federal addition, FDA may publish fullive and concerner of cation submission submission does not affect Register. Please note: under sections 531 through 542 of the Act for devices under the ally obligation you might have ander provisions, or other Federal laws or regulations. {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 nouncation. The I Dr I mailig of rassification 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 11 you desire specific acritic diagnostic devices), please contact the Office of Compliance at additionally 607.10 for in in in and advertising on the promotion and advertising of your device, (201) 594-1568. Frauncenf 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 information on your responsive Assistance at its toll-free number (800) 638-2041 or Manufacturers internet address "http://www.fda.gov/cdrl/dsma/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 {7}------------------------------------------------ ## Appendix G. Indications for Use Statement # INDICATIONS FOR USE STATEMENT 510(K) NUMBER : KOI2053 # DEVICE NAME : Is anti-Cardiolipin Screen Test System Indications for Use : The Diamedix Is anti-Cardiolipin Screen Test Kit is an indirect enzyme immunoassay (EIA) for the semi-quantitative measurement of IgG, IgM and IgA antibodies to cardiolipin in human serum as an aid in the assessment of the risk of thrombosis in patient with SLE or SLE-like disorders. These reagents can be used either manually or in conjunction with the MAGO® Plus Automated EIA Processor Josephine Bautista Division of Clinical Laboratory Devices 510(k) Number