BORRELIA BURGDORFERI IGG/IGM ELISA TEST SYSTEM

K033083 · Trinity Biotech USA · LSR · Nov 26, 2003 · Microbiology

Device Facts

• Record ID: K033083
• Device Name: BORRELIA BURGDORFERI IGG/IGM ELISA TEST SYSTEM
• Applicant: Trinity Biotech USA
• Product Code: LSR · Microbiology
• Decision Date: Nov 26, 2003
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3830
• Device Class: Class 2

Intended Use

The Trinity Biotech Captia™ Borrelia burgdorferi IgG/IgM ELISA kit is an Enzyme-Linked Immunosorbent Assay (ELISA) for the qualitative presumptive detection of total (IgG/IgM) antibodies to Borrelia burgdorferi in human serum. This ELISA should only be used for patients with signs and symptoms that are consistent with Lyme disease. Equivocal or positive results must be supplemented by testing with a standardized Western blot procedure. Positive supplemental results are supportive evidence of exposure to B. burgdorferi and can be used to support a clinical diagnosis of Lyme disease.

Device Story

ELISA kit for qualitative detection of total (IgG/IgM) antibodies to Borrelia burgdorferi in human serum; utilizes microassay plate coated with B. burgdorferi (strain B-31) antigen. Patient serum incubated in wells; antigen-specific antibodies bind to solid-phase antigen. After washing, goat anti-human IgG conjugated with horseradish peroxidase added; binds to antigen-antibody complexes. TMB substrate added; enzymatic reaction produces color proportional to antibody concentration. Reaction stopped with sulfuric acid; absorbance measured at 450 nm via spectrophotometer. Used in clinical laboratory settings by trained personnel. Results provide presumptive evidence of exposure; must be confirmed by standardized Western blot. Supports clinical diagnosis of Lyme disease when combined with patient history, signs, and symptoms.

Clinical Evidence

Performance evaluated using a CDC serum panel (n=47) and a clinical study of fresh patient sera (n=176). CDC panel results showed 71% agreement with clinical diagnosis. Clinical study compared device to BioWhittaker Lyme STAT; 1-step positive/equivocal results were 9.1% for Trinity vs 7.4% for predicate. Precision testing (n=810 determinations) showed inter-assay CVs generally <15% for most samples. Cross-reactivity testing performed against various conditions (lipemic, bilirubinemic, RPR+, RF+, EBV+, CMV+, RMS+, elevated ESR, CRP, dsDNA+).

Technological Characteristics

Enzyme-Linked Immunosorbent Assay (ELISA) utilizing purified B. burgdorferi antigen immobilized on solid-phase microtiter wells. Detection via enzyme-labeled anti-human IgG/IgM conjugate and substrate-induced colorimetric reaction. Photometric measurement of optical density. Manual or automated microplate reader required. No specific materials or connectivity standards provided.

Indications for Use

Indicated for qualitative presumptive detection of total (IgM/IgG) antibodies to Borrelia burgdorferi in human serum for patients with signs and symptoms consistent with Lyme disease. Equivocal or positive results require supplemental Western blot testing.

Regulatory Classification

Identification

Treponema pallidum treponemal test reagents are devices that consist of the antigens, antisera and all control reagents (standardized reagents with which test results are compared) which are derived from treponemal sources and that are used in the fluorescent treponemal antibody absorption test (FTA-ABS), the Treponema pallidum immobilization test (T.P.I.), and other treponemal tests used to identify antibodies to Treponema pallidum directly from infecting treponemal organisms in serum. The identification aids in the diagnosis of syphilis caused by bacteria belonging to the genus Treponema and provides epidemiological information on syphilis.

Predicate Devices

• Borrelia Burgdorferi IgG/IgM ELISA Test System (K965131)

Submission Summary (Full Text)

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K033083

# NOV 26 2003

Summary of Safety and Effectiveness Information Borrelia burgdorferi IgG/IgM ELISA Test Kit

I. Trinity Biotech 2823 Girts Road Jamestown, NY 14701 Contact person: Bonnie B. DeJoy Telephone: [phone] Date of preparation: Nov. 20, 2003

II. Description of Device

burgdorferi IgG/IgM ELISA kit is an The Borrelia Enzyme-Linked ImmunosorbentAssay (ELISA) for the qualitative presumptive detection of total (IgG/IgM) antibodies to Borrelia burgdorferi in human serum. This ELISA should only be used for patients with signs and patients with symptoms that are consistent with Lyme disease. Equivocal or positive results should be supplemented by testing with a standardized Western blot procedure. Positive supplemental results are supportive evidence of exposure to B. burgdorferi and can be used to support a clinical diagnosis of Lyme disease.

The Borrelia burgdorferi IgG/IgM ELISA test is an enzyme linked immunosorbent assav to detect IgG/IgM antibodies to Borrelia burgdorferi. Purified Borrelia burgdorferi antigen is attached to a solid phase microtiter well. Diluted test sera is added to each well. If the antibodies are present that recognize the antigen, they will bind to the antigen in the well. After incubation, the wells are washed to remove unbound antibody. An enzyme labeled anti-human IgG/IgM is added to each well. If antibody is present, it will bind to the antibody attached to the antigen on the well. After incubation the wells are washed to remove unbound conjugate. A substrate solution is added to each well. If enzyme is present. the substrate will undergo a color change. After an incubation period the reaction is stopped and the color intensity is measured photometrically, producing and indirect measurement of specific antibody in the patient specimen.

III. Predicate Device

The Borrelia burgdorferi IgG/IgM ELISA test is substantially equivalent to BioWhittaker's Lyme STAT test. Equivalence is demonstrated by the following comparative results:

### Performance Characteristics

### The CDC Lyme Disease Serum Panel Stratified by Time After Onset

The following information is from a serum panel obtained from the CDC and tested by Trinity Biotech. The results are presented as a means to convey further information on the performance of this assay with a masked, characterized serum panel. This does not imply an endorsement of the assay by the CDC.

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| Time From Onset | positive | equivocal | negative | Total | % Agreement<br>with Clinical<br>Diagnosis |
|-----------------|----------|-----------|----------|-------|-------------------------------------------|
| normals | 0 | 0 | 5 | 5 | 100% |
| < 1 month | 2 | 0 | 3 | 5 | 40.0% |
| 1-2 months | 6 | 2 | 2 | 10 | 80.0% |
| 3-12 months | 9 | 3 | 7 | 19 | 63.3% |
| > 1yr | 8 | 0 | 0 | 8 | 100.0% |
| Total | 25 | 5 | 17 | 47 | |

Table 1 Trinity Biotech B. burgdorferi IgG/IgM ELISA Results

Because the equivocals would have been tested by immunoblotting in a 2-step test system, they were considered 1-step positive for purposes of calculating % agreement. The Trinity Bitoech Borrelia burgdorferi IgG/IgM ELISA demonstrated 71% (30/42) agreement with clinical diagnosis of Lyme disease.

#### Study 2

One hundred seventy-six fresh sera from patients of various ages and genders that were submitted to a large clinical lab for B. burgdorferi antibody testing were tested on the Trinity Biotech B. burgdorferi IgG/IgM ELISA and BioWhittaker Lyme Stat. Any serum found positive or equivocal was tested by Mardx Diagnostics Western Blot on both IgG and IgM. The results are illustrated in Table 2.

#### Table 2

#### Western-blot

| Trinity<br>B. burgdorferi IgG/IgM | + | 6 | - | 3 |
|-----------------------------------|----|---|---|---|
| | eq | 1 | | 6 |
| Lyme Stat | + | 5 | - | 3 |
| | eq | 0 | | 5 |

| Type of Result (95%CI) | Trinity (95%CI) | Lyme Stat (95%CI) |
|------------------------------------------|---------------------------------|---------------------------------|
| 1-step (ELISA) Pos. or Eq. | 9.1% (4.8% - 13.4%)<br>(16/176) | 7.4% (3.4% - 11.3%)<br>(13/176) |
| 1-step Pos. or Eq. &<br>2-step (WB) Pos. | 4% (1.0% - 6.9%)<br>(7/176) | 2.8% (0.3% - 5.3%)<br>(5/176) |
| 2-step Pos. among<br>65.4%) | 44% (18.9% - 68.6%) | 39% (11.5% - ) |
| 1-step Pos. or Eq. | (7/16) | (5/13) |

CI = Confidence Interval

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#### Precision

Seven sera were assayed ten times each on three different plates at three different sites. An additional three sera were assayed ten times each on three different plates at two different sites. The intersite precision is shown in Table 3. With appropriate technique the user should obtain precision of <15% CV.

## Table 3 Trinity Biotech Borrelia burgdorferi IgG/IgM ELISA Inter-Assay Precision Between Sites

| Inter-Assay (n=90) | | | | | |
|--------------------|------|-------|--------|----|--|
| Serum # | X | SD | CV | n | |
| 1 | 1.18 | 0.113 | 9.58% | 90 | |
| 2 | 2.26 | 0.226 | 10.02% | 90 | |
| 3 | 3.85 | 0.278 | 7.21% | 90 | |
| 4 | 5.36 | 0.402 | 7.50% | 90 | |
| 5 | 2.04 | 0.186 | 9.11% | 90 | |
| 6 | 0.33 | 0.132 | 40.57% | 90 | |
| 7 | 0.34 | 0.234 | 68.93% | 90 | |
| 8 | 1.47 | 0.152 | 10.34% | 60 | |
| 9 | 1.47 | 0.104 | 7.10% | 60 | |
| 10 | 1.65 | 0.131 | 7.95% | 60 | |
| HPC | 5.31 | 0.314 | 5.91% | 15 | |
| LPC | 2.70 | 0.107 | 3.96% | 15 | |
| NC | 0.20 | 0.043 | 21.97% | 15 | |
| CAL | 3.34 | 0.103 | 3.08% | 45 | |

A total of 810 determinations were made at the three sites. In all 810 determinations there was

not one case of a positive result for a negative serum or a negative result for a positive serum.

#### X=Mean ISR

SD=Standard Deviation

CV=Coefficient of Variation=SD/X x 100

The methods in NCCLS EP5 were utilized for precision parameters.

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# Cross-Reactivity

The following potentially cross-reactive sera were run on the Trinity Biotech Borrelia burgdorferi IgG/IgM ELISA assay to assess cross-reactivity with the assay: lipemic, bilirubinemic, RPR+, dsDNA+, RF+, EBV+, CMV+, RMS+, elevated ESR, and CRP. The data in Table 4 illustrate the amount of reactivity with the sera.

# Table 4 Cross-Reactivity

| Laboratory result (Titer) positives | # of samples | # of |
|----------------------------------------------------|--------------|------|
| Lipemic (+ + +) | 5 | 0 |
| Bilirubinemic (1.9-16.9) | 5 | 1 |
| RPR + (1:2-1:64) | 10 | 0 |
| Rheumatoid Factor + (1:40-1:320) | 3 | 0 |
| Epstein Barr Virus Antibody + (1:40-1:2560) | 7 | 0 |
| Cytomegalovirus Antibody + (O.D. 0.718-2.308) | 6 | 1 |
| Rocky Mt Spotted Fever Antibody + (1:256-1:16,384) | 4 | 0 |
| CRP + (2.79-8.61 mg/dl) | 5 | 0 |
| Elevated ESR (40-115) | 10 | 0 |
| dsDNA + (52.3-1072 IU) | 16 | 0 |

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Image /page/4/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular border with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES • USA" arranged around the top half of the circle. Inside the circle is a stylized symbol that resembles three abstract human profiles facing right, with flowing lines suggesting movement or connection.

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

NOV 26 2003

Ms. Bonnie B. DeJoy Director, Quality Systems Trinity Biotech USA P.O. Box 1059 Jamestown, NY 14702-1059

Re: k033083

> Trade/Device Name: Captia Borrelia burgdorferi IgG/IgM ELISA Regulation Number: 21 CFR 866.3830 Regulation Name: Treponema pallidum treponemal test reagents Regulatory Class: Class II Product Code: LSR Dated: September 17, 2003 Received: September 29, 2003

Dear Ms. DeJoy:

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 Title 21, Code of Federal Regulations (CFR), Parts 800 to 895. 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 Parts 801 and 809); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820).

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Page 2 -

This letter will allow you to begin marketing your device as described in your Section 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 information about the application of labeling requirements to your device, or questions on the promotion and advertising of your device, please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at [phone]. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). You may obtain other general information on your responsibilities under the Act 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 Putman

Steven I. Gutman, M.D., M.B.A. Director Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

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Page 1 of 1

Over-The-Counter Use

510(k) Number: K033083

Device Name: Trinity Biotech Captia™ Borrelia burgdorferi IgG/IgM ELISA

Indications For Use: The Trinity Biotech Captia™ Borrelia burgdorferi IgG/IgM ELISA kit is an Enzyme-Linked Immunosorbent Assay (ELISA) for the qualitative presumptive detection of total (IgG/IgM) antibodies to Borrelia burgdorferi in human serum. This ELISA should only be used for patients with signs and symptoms that are consistent with Lyme disease. Equivocal or positive results must be supplemented by testing with a standardized Western blot procedure. Positive supplemental results are supportive evidence of exposure to B. burgdorferi and can be used to support a clinical diagnosis of Lyme disease.

# PLEASE DO NOT WRITE BELOW THIS LINE – CONTINUE ON ANQTHER PAGE IF NEEDED

Concurrence of CDRH, Office of Device Evaluation (ODE)

Prescription Use (Per 21 XFR 801.109) OR

(Optional Format 1-2-96)

Division Sign-Off for fMP

Office of In Vitro Diagnostic Device Evaluation and Safety

510(k) K033083