Lyme B. burgdorferi (IgG) MarStripe Test

K163095 · Trinity Biotech · LSR · Feb 1, 2017 · Microbiology

Device Facts

• Record ID: K163095
• Device Name: Lyme B. burgdorferi (IgG) MarStripe Test
• Applicant: Trinity Biotech
• Product Code: LSR · Microbiology
• Decision Date: Feb 1, 2017
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3830
• Device Class: Class 2

Intended Use

Lyme B. burgdorferi (IgG) MarStripe Test is an immunoblot assay for the in vitro qualitative detection of human IgG antibody to individual proteins of Borrelia burgdorferi in human serum or plasma (K₂-EDTA) in samples which have been found positive or equivocal using an EIA or IFA test procedure to provide supportive evidence of infection with B. burgdorferi.

Device Story

Lyme B. burgdorferi (IgG) MarStripe Test is an immunoblot assay for detecting IgG antibodies against B. burgdorferi antigens. Input: human serum or plasma (K2-EDTA) previously screened as positive/equivocal by EIA or IFA. Operation: patient sample incubated with nitrocellulose test strips containing purified B. burgdorferi antigens and control lines; reactive antibodies bind to antigens; anti-human IgG-HRP conjugate and TMB substrate added to produce visible blue/purple lines. Output: visual qualitative assessment of band intensity compared to a Cut-Off control line. Interpretation: strips with 5 or more of 10 test lines are positive for specific IgG antibody. Used in clinical laboratory settings by trained personnel. Results provide supportive evidence of B. burgdorferi infection, aiding clinical diagnosis of Lyme disease.

Clinical Evidence

Clinical study of 753 prospectively collected specimens compared the subject device to the predicate. Positive agreement was 92.9% (95% CI: 88.6%-95.7%) and negative agreement was 97.7% (95% CI: 95.9%-98.7%). Discrepant analysis using two additional Western blot methods confirmed the subject device's performance. Sensitivity was evaluated using 94 well-characterized clinical specimens (19.5% overall) and CDC reference panels (16.7% overall), showing performance comparable to the predicate. Analytical specificity was 99.5% (n=219). Interference and matrix equivalence (serum vs. plasma) studies showed 100% qualitative agreement.

Technological Characteristics

Immunoblot assay using nitrocellulose test strips with purified B. burgdorferi antigens and control lines (SAC, CAC, Cut-Off). Reagents include sample diluent (BSA/PBS), anti-human IgG-HRP conjugate, and TMB substrate. Qualitative visual readout. Manual test procedure. No software or electronic components.

Indications for Use

Indicated for the qualitative detection of human IgG antibodies to B. burgdorferi in human serum or plasma (K₂-EDTA) for patients previously testing positive or equivocal by EIA or IFA, to provide supportive evidence of Lyme disease infection. For prescription use only.

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

• MarDx B. burgdorferi IgG MarBlot Strip Test System (K950829)

Submission Summary (Full Text)

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Image /page/0/Picture/2 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a stylized caduceus symbol, which features three abstract human figures connected by a flowing line. The symbol is encircled by the text "DEPARTMENT OF HEALTH & HUMAN SERVICES • USA" in a circular arrangement.

February 1, 2017

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

Immco Diagnostics, Inc. Kevin Lawson Regulatory Officer 60 Pineview Dr. Buffalo, New York 14031

Re: K163095

Trade/Device Name: Lyme B. Burgdorferi (IgG) MarStripe Test Regulation Number: 21 CFR 866.3830 Regulation Name: Treponema pallidum treponemal test reagents Regulatory Class: Class II Product Code: LSR Dated: September 10, 2016 Received: November 4, 2016

Dear Kevin Lawson:

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. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to 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 and Part 809); medical device reporting (reporting of

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medical device-related adverse events) (21 CFR 803); 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.

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801 and Part 809), please contact the Division of Industry and Consumer Education at its toll-free number [phone] or [phone] or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to

http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number [phone] or [phone] or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely.

# Steven R. Gitterman -S

Uwe Scherf, Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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| DEPARTMENT OF HEALTH AND HUMAN SERVICES<br>Food and Drug Administration<br>Indications for Use | Form Approved: OMB No. 0910-0120<br>Expiration Date: January 31, 2017<br>See PRA Statement below. |
|------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| 510(k) Number (if known) | K163095 |
| Device Name | Lyme B. burgdorferi (IgG) MarStripe Test |
| Indications for Use (Describe) | Lyme B. burgdorferi (IgG) MarStripe Test is an immunoblot assay for the in vitro qualitative detection of human IgG antibody to individual proteins of <i>Borrelia burgdorferi</i> in human serum or plasma (K2- EDTA) in samples which have been found positive or equivocal using an EIA or IFA test procedure to provide supportive evidence of infection with <i>B. burgdorferi</i> . |

Type of Use (Select one or both, as applicable)

| <div> <span style="text-decoration: underline;">☑</span> Prescription Use (Part 21 CFR 801 Subpart D) </div> | <div> ☐ Over-The-Counter Use (21 CFR 801 Subpart C) </div> |
|--------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------|
|--------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------|

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Image /page/3/Picture/0 description: The image shows the logo for Immco Diagnostics, a Trinity Biotech Company. The logo features a cluster of blue and green dots on the left, followed by the company name "immco" in blue, and "DIAGNOSTICS" in green below it. Underneath the company name, it says "A Trinity Biotech Company" in a smaller font.

# 510(k) Summary of Safety and Effectiveness

This 510(k) summary of safety and effectiveness information is being submitted in accordance with the requirement of SMDA 1990 and 21 CFR 807.92.

| 1. Submitter Name: | Immco Diagnostics, Inc. a Trinity Biotech Company |
|--------------------|---------------------------------------------------|
| Address: | 60 Pineview Dr., Buffalo, NY 14228 |
| Phone Number: | 716-691-0091 ext. 110 |
| Contact Person: | Kevin Lawson |
| Summary Prepared: | 1-4-2017 |

- Lyme B. burgdorferi (IgG) MarStripe Test 2. Device Name: Common Name: Lyme B. burgdorferi (IgG) Immunoblot Product Code: LSR
- 3. Substantially equivalent to: MarDx B. burgdorferi IgG MarBlot Strip Test System (K950829)
- 4. Description of device: The kit is an immunoblot method to detect IgG antibodies against B. burqdorferi antigens. The test kit contains:
- . Nitrocellulose Test Strips with purified B. burgdorferi antigens (10) and quality control lines (3) present in specific positions
- . Sample Diluent. Provided for specimen dilutions. Contains BSA and PBS
- Positive Control derived from human serum positive for Lyme disease. Contains <0.1% sodium azide
- Negative Control derived from human serum negative for Lyme disease. Contains <0.1% sodium azide
- . Conjugate. Antihuman IgG-HRP Conjugate binds reactive antibodies to the Substrate
- TMB Substrate. Provides colorimetric reaction for visual read of bound antibodies
- PBS Wash Buffer concentrate. Removes reagents and unbound antibodies after incubation steps. Must be reconstituted to 1L with distilled or deionized water.

To perform the test, serum or plasma is individual B. burgdorferi Test Strips. In positive sera antibodies specifically bind to one or more of the test lines on the strips are washed according to the protocol, and then the pre-diuted, readyto-use Conjugate is added to the test strips. After incubation and wash steps, the ready-to-use Substrate is added to the strips. During a 10 minute (±4 min) incubation, conjugate and substrate binding produces visible blue/purple lines for Serum Addition Control (SAC), Conjugate Addition Control (CAC) and Cut-Off Control lines. If the sample is positive for any of the antigen coated test lines, it will show a reaction more intense than the Cut-Off line. Reactions are read visually and reported as positive or equivocal (comparable to Cut-Off line). Strips which have 5 (or more) of the 10 test lines are considered positive for specific IgG antibody to B. burgdorferi.

- Intended Use: Lyme B. burqdorferi (IgG) MarStripe Test is an immunoblot assay for the in vitro qualitative detection of human 5. lgG antibody to individual proteins of Borrelia burgdorferi in human serum or plasma (K- EDTA) in samples which have been found positive or equivocal using an EIA or IFA test procedure to provide supportive evidence of infection with B. burgdorferi.
- 6. Similarities and Differences: The Lyme B. burgdorferi (IgG) MarStripe Test was compared to a commercially marketed kit by Trinity Biotech the B. burgdorferi (lgG) MarBlot Strip Test System (K950829). Both kits have the same intended use and use the same methodology except that the MarStripe Test has been validated in plasma (K- EDTA) as well as serum. Both immunoblot kits detect antibodies to B. burgdorferi antigens using the standard immunoblot IgG algorithm. Both assays are qualitative. The MarStripe uses a Horseradish Peroxidase Conjugate and Tetramethylbenzidine (TMB) Substrate/Chromogen in comparison to The MarBlot Alkaline Phosphatase Conjugate and BCIP/NBT Substrate/Chromogen. The MarStripe uses a Cutoff Control line incorporated in the strip in comparison to the 41kD band of Weakly Reactive Control.

## 7. Non-clinical Tests:

60 Pineview Drive Buffalo, NY 14228-2120 USA Toll free [phone]  •  tel. [phone]  •  fax [phone]

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Image /page/4/Picture/0 description: The image shows the logo for Immco Diagnostics, a Trinity Biotech Company. The logo features a cluster of blue and green circles on the left, followed by the word "immco" in blue, bold letters. Below "immco" is the word "DIAGNOSTICS" in smaller, green letters, and below that is the phrase "A Trinity Biotech Company" in a smaller, lighter font.

## Reproducibility

Eight specimens were tested by Lyme B. burgdorferi (lgG) MarStripe Test in 4 replicates, two runs per days for a total of 72 tests for each specimen at three laboratory sites. Results were read by 2 human operators at each site, equaling a total of 432 read-outs. Samples were selected based on FDA cleared B. burgdorferi ELISA results, including 2 low negative samples, 2 high negative samples, 2 low positive samples. Final positive samples. Final positive or negative agreement was 100% for all specimens.

| Sample | | p93 | p66 | p58 | p45 | p41 | p39 | p30 | p28 | p23 | p18 |
|--------|--------------|-------|------|------|------|------|------|------|------|------|------|
| 1 | n=432 | | | | | | | | | | |
| | Low Neg | Neg. | Neg. | Neg. | Neg. | Neg. | Neg. | Neg. | Neg. | Neg. | Neg. |
| | Band Type | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| | Positives | 432 | 432 | 432 | 432 | 432 | 432 | 432 | 432 | 432 | 432 |
| | Negatives | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% |
| 2 | % Positive | | | | | | | | | | |
| | Low Neg | Neg. | Neg. | Neg. | Neg. | Pos. | Neg. | Neg. | Neg. | Neg. | Neg. |
| | Band Type | 0 | 0 | 0 | 0 | 432 | 0 | 0 | 0 | 0 | 0 |
| | Positives | 432 | 432 | 432 | 432 | 0 | 432 | 432 | 432 | 432 | 432 |
| | Negatives | 0% | 0% | 0% | 0% | 100% | 0% | 0% | 0% | 0% | 0% |
| 3 | % Positive | | | | | | | | | | |
| | High Neg | Neg. | Neg. | Neg. | Neg. | Neg. | Neg. | Neg. | Neg. | Neg. | Neg. |
| | Band Type | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| | Positives | 432 | 432 | 432 | 432 | 432 | 432 | 432 | 432 | 432 | 432 |
| | Negatives | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% |
| 4 | % Positive | | | | | | | | | | |
| | High Neg | Neg. | Neg. | Neg. | Neg. | Pos. | Neg. | Neg. | Neg. | Neg. | Neg. |
| | Band Type | 0 | 0 | 0 | 0 | 432 | 0 | 0 | 0 | 0 | 0 |
| | Positives | 432 | 432 | 432 | 432 | 0 | 432 | 432 | 432 | 432 | 432 |
| | Negatives | 0% | 0% | 0% | 0% | 100% | 0% | 0% | 0% | 0% | 0% |
| 5 | % Positive | | | | | | | | | | |
| | Low Pos | Neg. | Neg. | Neg. | Neg. | Pos. | Neg. | Neg. | Neg. | Pos. | Neg. |
| | Band Type | 0 | 0 | 0 | 0 | 432 | 0 | 0 | 0 | 432 | 0 |
| | Positives | 432 | 432 | 432 | 432 | 0 | 432 | 432 | 432 | 0 | 432 |
| | Negatives | 0% | 0% | 0% | 0% | 100% | 0% | 0% | 0% | 100% | 0% |
| 6 | % Positive | | | | | | | | | | |
| | Low Pos | Neg. | Neg. | Neg. | Neg. | Pos. | Neg. | Neg. | Neg. | Pos. | Neg. |
| | Band Type | 0 | 0 | 0 | 0 | 432 | 0 | 0 | 0 | 432 | 0 |
| | Positives | 432 | 432 | 432 | 432 | 0 | 432 | 432 | 432 | 0 | 432 |
| | Negatives | 0% | 0% | 0% | 0% | 100% | 0% | 0% | 0% | 100% | 0% |
| 7 | % Positive | | | | | | | | | | |
| | Moderate Pos | Wpos. | Pos. | Pos. | Neg. | Pos. | Pos. | Cut. | Pos. | Pos. | Pos. |
| | Band Type | 412 | 432 | 432 | 0 | 432 | 432 | 412 | 432 | 432 | 432 |
| | Positives | 20 | 0 | 0 | 432 | 0 | 0 | 20 | 0 | 0 | 0 |
| | Negatives | 95% | 100% | 100% | 0% | 100% | 100% | 95% | 100% | 100% | 100% |
| 8 | % Positive | | | | | | | | | | |
| | Moderate Pos | Pos. | Neg. | Pos. | Neg. | Pos. | Pos. | Neg. | Neg. | Pos. | Neg. |
| | Band Type | 432 | 0 | 432 | 0 | 432 | 432 | 0 | 0 | 432 | 0 |
| | Positives | 0 | 432 | 0 | 432 | 0 | 0 | 432 | 432 | 0 | 432 |
| | Negatives | 100% | 0% | 100% | 0% | 100% | 100% | 0% | 0% | 100% | 0% |
| | % Positive | | | | | | | | | | |

Pos = positive band. Neg = negative band. Wpos = weak positive band. Cut = equivocal band.

Analytical Specificity: 219 sera from healthy blood donors representing endemic geographic regions of the United States were tested. Analytical specificity was determined to be 99.5% (95% Cl: 97.1% - 100%).

| | | Normal Individuals |
|-------------------------------|----------|--------------------|
| Lyme IgG<br>MarStripe<br>Test | Positive | 1 |
| | Negative | 218 |
| | Total | 219 |

60 Pineview Drive ● ● Buffalo, NY 14228-2120 ● ● USA Toll free [phone]  •  tel. [phone]  •  fax [phone]

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Image /page/5/Picture/0 description: The image shows the logo for Immco Diagnostics. The logo features a cluster of blue and green dots on the left, followed by the company name in blue and green text. Below the company name, it says "A Trinity Biotech Company" in smaller, gray text.

Cross-reactivity: A total of 136 potentially cross-reactive specimens from individuals with other or infectious conditions were tested on this assay. Positive incidence was 4.4% and all positives specimens by the Lyme B. burgdorferi (IgG) MarStripe Test were confirmed positive when tested by the predicate IgG Western blot device.

| Population | n | n Positive | % Positive |
|------------------------------|-----|------------|------------|
| Ehrlichia chafeensis | 10 | 0 | 0 |
| Babesia microti | 10 | 4 | 40 |
| Leptospira interrogans | 10 | 0 | 0 |
| Helicobacter pylori | 10 | 2 | 20 |
| Syphilis | 10 | 0 | 0 |
| Influenza | 10 | 0 | 0 |
| Rocky Mountain Spotted Fever | 10 | 0 | 0 |
| Parvovirus B19 | 9 | 0 | 0 |
| Systemic lupus erythematosus | 15 | 0 | 0 |
| Cytomegalovirus | 10 | 0 | 0 |
| Rheumatoid arthritis | 16 | 0 | 0 |
| Celiac | 16 | 0 | 0 |
| Total | 136 | 6 | 4.4 |

Interference: Two Lyme IgG negative and three Lyne IgG positive sera were spiked with hemoglobin (2g/L), unconjugated bilirubin (342 µmol/L), RF (100 IU/ml), triglycerides (3.7 mmol/L) and total cholesterol (13 mmol/L) and tested using this assay. Samples were tested with and without interfering agents. Qualitative agreement was 100% for all specimens.

Serum vs. Plasma Matrix Comparison Studies: To establish equivalence of serum vs. plasma (Kg-EDTA) matrix, 10 pairs of sera/plasma (samples A-J below) were sourced from specimens tested on an FDA cleared Lyme EIA assay. Two of these were Western blot lgG positive, 8 were negative. These specimens were selectively pooled to create another set of 10 samples (samples 1-10 below), including 3 Western blot IgG positives. These samples were assyed on the Lyme B. burgdorferi (IgG) MarStripe Test. Qualitative agreement for all pairs was 100%.

Image /page/5/Picture/6 description: The image displays the text "www.immco.com" in a blue, sans-serif font. The text is underlined with a solid blue line. The background of the image is plain white, providing a clean and simple presentation of the website address.

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Image /page/6/Picture/0 description: The image shows the logo for Immco Diagnostics. The logo features a cluster of blue and green circles to the left of the word "immco" in blue, with a small blue circle above the "o". Below "immco" is the word "DIAGNOSTICS" in green. Underneath that, it says "A Trinity Biotech Company" in a smaller font.

| Sample ID1 | Type2 | p93 | p66 | p58 | p45 | p41 | p39 | p30 | p28 | p23 | p18 | Result | Band % Pos<br>Agrmt | Band % Neg<br>Agrmt |
|---------------|--------|-----|-----|-----|-----|-----|-----|-----|-----|-----|-----|--------|---------------------|---------------------|
| A | Serum | 0 | 0 | 0 | 0 | 1 | 0 | 0 | 0 | 1 | 0 | NEG | | 100 |
| A | Plasma | 0 | 0 | 0 | 0 | 1 | 0 | 0 | 0 | 1 | 0 | NEG | 100 | |
| B | Serum | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | | 100 |
| B | Plasma | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | 100 | |
| C | Serum | 1 | 0 | 1 | 0 | 1 | 1 | 1 | 0 | 1 | 1 | POS | | 100 |
| C | Plasma | 1 | 0 | 1 | 0 | 1 | 1 | 1 | 0 | 1 | 1 | POS | 100 | |
| D | Serum | 0 | 0 | 1 | 0 | 1 | 1 | 1 | 0 | 1 | 1 | POS | | 100 |
| D | Plasma | 0 | 0 | 1 | 0 | 1 | 1 | 1 | 0 | 1 | 1 | POS | 100 | |
| E | Serum | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | NEG | 100 | |
| E | Plasma | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | NEG | | 100 |
| F | Serum | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | 100 | 100 |
| F | Plasma | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | | |
| G | Serum | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | 100 | 100 |
| G | Plasma | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | | |
| H | Serum | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | 100 | 100 |
| H | Plasma | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | | |
| I | Serum | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | 100 | 100 |
| I | Plasma | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | | |
| J | Serum | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | 100 | 100 |
| J | Plasma | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | | |
| Pool 1 (A+B) | Serum | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | NEG | 100 | 100 |
| Pool 1 (A+B) | Plasma | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | NEG | | |
| Pool 2 (B+C) | Serum | 1 | 0 | 1 | 0 | 1 | 1 | 0 | 0 | 1 | 1 | POS | 100 | 100 |
| Pool 2 (B+C) | Plasma | 1 | 0 | 1 | 0 | 1 | 1 | 0 | 0 | 1 | 1 | POS | | |
| Pool 3 (C+D) | Serum | 1 | 0 | 1 | 0 | 1 | 1 | 1 | 0 | 1 | 1 | POS | 100 | 100 |
| Pool 3 (C+D) | Plasma | 1 | 0 | 1 | 0 | 1 | 1 | 1 | 0 | 1 | 1 | POS | | |
| Pool 4 (D+E) | Serum | 0 | 0 | 1 | 0 | 1 | 1 | 0 | 0 | 1 | 1 | POS | 100 | 100 |
| Pool 4 (D+E) | Plasma | 0 | 0 | 1 | 0 | 1 | 1 | 0 | 0 | 1 | 1 | POS | | |
| Pool 5 (A+E) | Serum | 0 | 0 | 0 | 0 | 1 | 0 | 0 | 0 | 1 | 0 | NEG | 100 | 100 |
| Pool 5 (A+E) | Plasma | 0 | 0 | 0 | 0 | 1 | 0 | 0 | 0 | 1 | 0 | NEG | | |
| Pool 6 (F+G) | Serum | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | 100 | |
| Pool 6 (F+G) | Plasma | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | | 100 |
| Pool 7 (G+H) | Serum | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | 100 | 100 |
| Pool 7 (G+H) | Plasma | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | | |
| Pool 8 (H+I) | Serum | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | 100 | 100 |
| Pool 8 (H+I) | Plasma | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | | |
| Pool 9 (I+J) | Serum | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | 100 | 100 |
| Pool 9 (I+J) | Plasma | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | | |
| Pool 10 (F+J) | Serum | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | 100 | 100 |
| Pool 10 (F+J) | Plasma | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | NEG | | |

1. Pool 1 to 10 contain a 1:1 mixture of the native samples as indicated. 2. 0 = negative band result. 1 = positive band result.

## 8. Clinical Tests:

Method Comparison: Three sites tested utilized prospectively collected consecutive specimens testing positive on a FDA cleared first-step ElA in the study. 753 specimens were tested on the predicate MarDx B. burgdorferi lgG MarBlot Test device as well as the Lyme B. burgdorferi (IgG) MarStripe Test

| | | Predicate IgG WB | | |
|----------------------------|----------|------------------|----------|-------|
| Lyme IgG<br>MarStripe Test | | Positive | Negative | Total |
| | Positive | 221 | 12 | 233 |
| | Negative | 17 | 503 | 520 |
| | Total | 238 | 515 | 753 |

Positive Agreement: 92.9% (95% CI: 88.6% - 95.7%) Negative Agreement: 97.7% (95% CI: 95.9% - 98.7%)

> 60 Pineview Drive ● Buffalo, NY 14228-2120 ● Toll free [phone]  •  tel. [phone]  •  fax [phone]

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Image /page/7/Picture/0 description: The image shows the logo for Immco Diagnostics. The logo features a series of blue and green dots on the left side, followed by the word "immco" in blue, with the word "DIAGNOSTICS" in green underneath. Below that, it says "A Trinity Biotech Company" in a smaller font.

The 29 discrepant specimens were tested on two additional commercially available B. burgdorferi IgG Western blot methods. Results of the Lyme B. burgdorferi (IgG) MarStripe Test were compared to the consensus (2/3 comparator) results. The 12 positives by MarStripe remained positive by consensus testing and the 17 negatives by MarStripe were found to be positives.

| | | Consensus Result | |
|----------------------------|----------|------------------|----------|
| | | Positive | Negative |
| Lyme IgG<br>MarStripe Test | Positive | 12 | 0 |
| | Negative | 17 | 0 |

Sensitivity: 94 well characterized Lyme disease clinical specimens from the above study were B. burgdorferi (IgG) MarStripe Test. Specimens included samples from early, early disseminated, and late phase. The sensitivity obtained was compared with that of the predicate device.

| Interval | n | Lyme IgG MarStripe<br>Test | | Predicate IgG WB | |
|---------------------------------|----|----------------------------|------|------------------|------|
| | | positive | % | positive | % |
| Early Lyme (stage 1) | 22 | 4 | 18.2 | 3 | 13.6 |
| Early disseminated<br>(stage 2) | 44 | 12 | 27.3 | 11 | 25.0 |
| Late Lyme (stage 3) | 28 | 5 | 17.9 | 5 | 17.9 |
| Overall | 94 | 21 | 19.5 | 19 | 20.2 |

Sensitivity Comparison:

Lyme B. burgdorferi (IgG) MarStripe Test: Predicate device: Difference in proportion:

22.3% (21/94) (95% Cl: 14.7%-32.3%) 20.2% (19/94) (95% Cl: 12.9%-30.0%) 2.1%

CDC Panels: Reference panels from the Center for Disease Control and Prevention (Lyme Disease Validation Panel n=10, Lyme Disease Basic Research Panel n=32) were tested on the Lyme B. burgdorferi (lgG) MarStripe Test and the predicate device.

| Interval | n | Lyme IgG<br>MarStripe Test | | Predicate IgG WB | |
|------------------------------|----|----------------------------|------|------------------|------|
| | | positive | % | positive | % |
| Controls | 25 | 0 | 0 | 0 | 0 |
| Early Lyme (stage 1) | 10 | 1 | 10 | 1 | 10 |
| Early disseminated (stage 2) | 3 | 3 | 100 | 3 | 100 |
| Late Lyme (stage 3) | 4 | 4 | 100 | 4 | 100 |
| Overall | 42 | 7 | 16.7 | 7 | 16.7 |

Note: 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.

- 9. Conclusion: From the performance data and kit comparison above, it is our conclusion that the Lyme B. burgdorferi (IgG) MarStripe Test is substantially equivalent to the B. burgdorferi (IgG) MarBlot Strip Test System (K950829) commercially marketed by Trinity Biotech
Klaus

Kevin J. Lawson VP Regulatory Affairs

60 Pineview Drive Buffalo, NY 14228-2120 537-8378 · fax [phone]