BORRELIA BURGDORFERI IGM ELISA TEST SYSTEM

K033070 · Trinity Biotech USA · LSR · Nov 26, 2003 · Microbiology

Device Facts

• Record ID: K033070
• Device Name: BORRELIA BURGDORFERI 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 IgM ELISA kit is an Enzyme-Linked Immunosorbent Assay (ELISA) for the qualitative presumptive detection of IgM antibodies to Borrelia burgdorferi in human serum. This ELISA should 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 presumptive detection of IgM antibodies to Borrelia burgdorferi in human serum; utilizes microassay plate coated with purified B. burgdorferi antigen (strain B-31). Patient serum incubated on plate; antigen-specific antibodies bind to solid-phase antigen. After washing, goat anti-human IgM conjugated with horseradish peroxidase added; binds to antigen-antibody complexes. Chromogen/substrate (TMB) added; enzymatic reaction produces color change proportional to antibody presence. Reaction stopped with sulfuric acid; optical density measured at 450 nm using microplate reader. Results interpreted via cutoff calibrator and correction factor. Used in clinical laboratory settings by trained personnel. Output supports clinical diagnosis of Lyme disease when combined with patient history, signs, symptoms, and mandatory supplemental Western blot testing.

Clinical Evidence

No clinical data provided. Performance characteristics were established in the predicate device (K965129).

Technological Characteristics

Enzyme-Linked Immunosorbent Assay (ELISA) utilizing purified B. burgdorferi antigen coated on solid-phase microtiter wells. Detection via enzyme-labeled anti-human IgM conjugate and photometric measurement of substrate color change. Manual or automated laboratory processing. No specific materials or software algorithms described beyond standard ELISA biochemical principles.

Indications for Use

Indicated for qualitative presumptive detection of IgM 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 IgM ELISA Test System (K965129)

Submission Summary (Full Text)

{0}------------------------------------------------

K033070

NOV 26 2003

Summary of Safety and Effectiveness Information Borrelia burgdorferi 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

The Borrelia burgdorferi IgM ELISA kit is an Enzyme-Linked Immunosorbent Assay (ELISA) for the qualitative presumptive detection of 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.

The Borrelia burgdorferi IgM ELISA test is an enzyme linked immunosorbent assay to detect IgM antibodies to Borrelia burgdorferi. Purified Borrelia burgdorferi antigen is attached to a solid phase microtiter well. Pretreated 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 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 an indirect measurement of specific antibody in the patient specimen.

#### III. Predicate Device

The Borrelia burgdorferi IgM ELISA test is substantially equivalent to Bio Whittaker's Lyme M STAT test. Equivalence is demonstrated by the following comparative results:

## Performance Characteristics

The CDC Lyme Disease Serum Panel Stratified by Time After OnsetThe 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.

{1}------------------------------------------------

| Time From<br>Onset | positive | equivocal | negative | Total | %Agreement<br>with Clinical<br>Diagnosis |
|--------------------|----------|-----------|----------|-------|------------------------------------------|
| normals | 0 | 0 | 5 | 5 | 100.0% |
| < 1 month | 3 | 1 | 1 | 5 | 80.0% |
| 1-2 months | 6 | 0 | 4 | 10 | 60.0% |
| 3-12 months | 7 | 2 | 10 | 19 | 47.4% |
| > 1 year | 1 | 0 | 7 | 8 | 12.5% |
| Total | 17 | 3 | 27 | 47 | |

Table 1 Trinity Biotech B. burgdorferi 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 Biotech B. burgdorferi IgM ELISA demonstrated 47.6% (20/42) agreement with clinical diagnosis of Lyme disease.

## Study 2

One hundred seventy-six 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 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 Biotech | + | | 6 | 10 | | |
| <i>B. burgdorferi</i> eq.<br>IgM | | | 0 | 3 | | |
| LYME STAT | + | | 6 | 9 | | |
| | eq. | | 0 | 3 | | |
| Type of Result | | | Trinity | (95%CI) | LYME STAT | (95%CI) |
| 1-step (ELISA) Pos. or Eq. | | | 10.8%<br>(19/176) | (6.1-15.5%) | 10.2%<br>(18/176) | (5.7-14.8%) |
| 1-step Pos. or Eq. &<br>2-step (WB) pos. | | | 3.41%<br>(6/176) | (0.7-6.1%) | 3.41%<br>(6/176) | (0.7-6.1%) |
| 2-step Pos. among<br>1-step Pos. or Eq. | | | 31.6%<br>(6/19) | (10.3-52.9%) | 33.3%<br>(6/18) | (11.1-55.6%) |
| CI = Confidence Interval | | | | | | |

{2}------------------------------------------------

### Precision

Seven sera were assayed ten times each on three different assays at two different sites. The intersite precision is shown in Table 3. With appropriate technique the user should obtain precision of <20% CV for positive samples, Calibrators and Positive Controls.

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

Inter-Assay (n=60)

| Serum # | X | SD | CV |
|---------|------|-------|--------|
| 1 | 3.74 | 0.335 | 8.96% |
| 2 | 1.95 | 0.188 | 9.64% |
| 3 | 0.94 | 0.105 | 11.2% |
| 4 | 0.05 | 0.034 | 68.0% |
| 5 | 0.98 | 0.155 | 15.82% |
| 6 | 1.34 | 0.187 | 13.96% |
| 7 | 1.24 | 0.213 | 17.18% |
| HPC * | 4.07 | 0.287 | 7.04% |
| Cal ** | 2.22 | 0.104 | 4.70% |
| LPC * | 2.21 | 0.113 | 5.11% |
| NC * | 0.05 | 0.031 | 64.0% |

$$\text{* For HPC, LPC, and NC } \text{ n} = 12.$$

** For Cal n = 36

A total of four hundred ninety-two determinations were made at the two sites. In all of the determinations there was not one case of a positive result for a negative serum or a negative result for a positive serum.

Equivocal sample #3 was negative 23 times and positive 19 times. Equivocal sample #5 was negative 19 times and positive 3 times. X=Mean ISR SD=Standard Deviation CV=Coefficient of Variation = $SD/X$ x 100

The methods in NCCLS EPS were utililized for precision parameters.

#### Cross-Reactivity

The following potentially cross-reactive sera were run on the Trinity Biotech Borrelia burgdorferi 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.

{3}------------------------------------------------

# Table 4 Cross-Reactivity

.

:

、

| Laboratory Result (Titer) | # of samples | # of positives |
|----------------------------------------------------|--------------|----------------|
| Lipemic (+++) | 5 | 0 |
| Bilirubinemic (1.9-16.9) | 5 | 0 |
| RPR + (1:2-1:64) | 10 | 0 |
| Rheumatoid Factor + (1:40-1:320) | 8 | 0 |
| Epstein Barr Virus Antibody +(1:40-1:2560) | 7 | 0 |
| Cytomegalovirus Antibody + (O.D. 0.718-2.308) | 6 | 0 |
| 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 |

{4}------------------------------------------------

Image /page/4/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized eagle with three lines forming its body and head. 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

NOV 26 2003

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

Re: k033070 Trade/Device Name: Captia Borrelia burgdorferi 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).

{5}------------------------------------------------

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

{6}------------------------------------------------

### Page 1 of 1

510(k) Number: K033070

Device Name: Trinity Biotech Captia™ Borrelia burgdorferi IgM ELISA

Indications For Use: The Trinity Biotech Captia™ Borrelia burgdorferi IgM ELISA kit is an Enzyme-Linked Immunosorbent Assay (ELISA) for the qualitative presumptive detection of IgM antibodies to Borrelia burgdorferi in human serum. This ELISA should 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 ANOTHER PAGE IF NEEDED

Concurrence of CDRH, Office of Device Evaluation (ODE)

| Prescription Use<br>(Per 21 XFR 801.109) | OR | Over-The-Counter Use<br>(Optional Format 1-2-96) |
|------------------------------------------|----|--------------------------------------------------|
|------------------------------------------|----|--------------------------------------------------|

| <div align="center">Division Sign-Off</div> |
|---------------------------------------------|

Office of In Vitro Diagnostic DeviceOmoo tion and Safety

510(k) Ro33070