Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELISA Test Kit

K180264 · Gold Standard Diagnostics · LSR · May 2, 2018 · Microbiology

Device Facts

• Record ID: K180264
• Device Name: Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELISA Test Kit
• Applicant: Gold Standard Diagnostics
• Product Code: LSR · Microbiology
• Decision Date: May 2, 2018
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3830
• Device Class: Class 2

Intended Use

The Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELISA Test kit is intended as a qualitative presumptive (first step) test for the detection of IgG and IgM antibodies to B. burgdorferi sensu stricto in human serum from symptomatic patient or people suspected of infection. Positive and equivocal results must be supplemented by testing with a second-step Western blot assay.

Device Story

The Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELISA Test Kit is an in vitro diagnostic assay used in clinical laboratories. It detects IgG and IgM antibodies to B. burgdorferi sensu stricto in human serum samples. The device utilizes an antigen-coated microtiter plate (B. burgdorferi B31 lysate, 2591 lysate, and recombinant VlsE) to capture specific antibodies. Following sample incubation and washing, a horseradish peroxidase-conjugated goat anti-human IgG/IgM antibody is added, followed by a TMB chromogenic substrate. The enzymatic reaction produces a color change (blue to yellow upon adding stop solution), which is measured photometrically at 450nm. Results are interpreted as positive, equivocal, or negative based on unit conversion. The test serves as a first-step presumptive screen; positive or equivocal results require confirmation via a second-step Western blot assay. The output assists clinicians in the diagnostic workup of patients suspected of having Lyme disease.

Clinical Evidence

Clinical sensitivity evaluated on 89 characterized Lyme disease samples (early, disseminated, late stages) and a CDC-provided panel. Sensitivity: 76.3% (early), 100% (disseminated), 97.2% (late). CDC panel agreement: 86.7% (early), 53.8% (convalescent), 100% (late). Method comparison with predicate (n=520) showed 96.0% positive percent agreement and 97.5% negative percent agreement. Analytical specificity (n=210) was 97.3% (endemic) and 98.0% (non-endemic).

Technological Characteristics

ELISA-based immunoassay. Components: polystyrene 96-well microtiter plates, B. burgdorferi antigens (B31, 2591, recombinant VlsE), goat anti-human IgG/IgM-HRP conjugate, TMB substrate, stop solution. Detection via photometric microplate reader at 450nm. Manual or automated processing. No specific software algorithm class stated; relies on standard photometric absorbance measurement.

Indications for Use

Indicated for symptomatic patients or those suspected of Lyme disease infection. For prescription use only. Positive/equivocal results require second-step Western blot confirmation.

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

• Trinity Biotech Captia™ Borrelia burgdorferi IgG/IgM ELISA Test Kit (K033070)

Submission Summary (Full Text)

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Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

May 2, 2018

Gold Standard Diagnostics Napoleon Monce Director, Product Development 2851 Spafford St. Davis, California 95618

Re: K180264

Trade/Device Name: Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELISA Test Kit Regulation Number: 21 CFR 866.3830 Regulation Name: Treponema pallidum treponemal test reagents Regulatory Class: Class II Product Code: LSR Dated: January 26, 2018 Received: January 30, 2018

Dear Napoleon Monce:

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 of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR

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Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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.

For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (http://www.fda.gov/DICE) for more information or contact DICE by email ([e-mail]) or phone [phone] or 301-796-7100).

Sincerely,

# Steven R. Gitterman -S

for

Uwe Scherf, M.Sc., 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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# Indications for Use

510(k) Number (if known) K180264

#### Device Name

Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELISA Test Kit

Indications for Use (Describe)

The Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELISA Test Kit is intended as a qualitative presumptive (first-step) test for the detection of IgG and IgM antibodies to B. burgdorferi sensu stricto in human serum from symptomatic patients or people suspected of infection. Positive and equivocal results must be supplemented by testing with a second-step Western blot assay.

| Type of Use (Select one or both, as applicable) | |
|----------------------------------------------------------------------------|---------------------------------------------------------------------------|
| <div> <span>☑</span> Prescription Use (Part 21 CFR 801 Subpart D) </div> | <div> <span>☐</span> Over-The-Counter Use (21 CFR 801 Subpart C) </div> |

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Image /page/3/Picture/0 description: The image is a logo for Gold Standard Diagnostics. The logo features the words "GOLD STANDARD" in large, bold, black letters. The word "GSD" is inside of a golden globe to the left of the word "GOLD". Below the words "GOLD STANDARD" is the word "DIAGNOSTICS" in smaller, black letters.

# 510(k) Summary

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

- 1) Submitter's Name: Gold Standard Diagnostics Address: 2851 Spafford St. Davis, CA. 95618 Phone Number: 530-759-8000 Contact Person: Napoleon Monce January 26, 2018 Date:
- 2) Product and Trade Name: Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELISA Test Kit

Common Name: Lyme ELISA Test

Regulation Section: (21 CFR 866.3830) Treponema pallidum treponemal test reagents.

Classification: Class II

Product Code: LSR; Reagent, Borrelia Serological Reagent

- 3) Legally Marketed Device to Which the Submitter Claims Equivalence:
Trinity Biotech Captia™ Borrelia burgdorferi IgG/IgM ELISA Test Kit (K033070).

## 4) Description of the Device:

The kit includes 12 x 8 well Antigen Coated strips, Conjugate, Substrate, Stop Solution, Wash Buffer. Diluent. Negative Control. Positive Control. The control. The controls are provided to determine if the assay is functioning properly and to determine the antibody level. The reagents are sufficient for 96 determinations.

During the test procedure, antibodies to B. burgdorferi (sensu stricto) if present in the human serum sample will bind to the antigens coated onto the wells forming antigen-antibody complexes. Excess antibodies are removed by washing. A conjugate of goat anti-human IgG/IgM antibodies conjugated with horseradish peroxidase are then added, which binds to the antigen-antibody complexes. Excess conjugate is removed by washing. This is followed by the addition of a chromogenic substrate, tetramethylbenzidine (TMB). If specific antibodies to the antigen are present in the patients' serum, a blue color will develop. The enzymatic reaction is then stopped with a stopping solution causing the contents of the well to turn yellow. The wells are read photometrically with a microplate reader at 450nm.

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The antigens used in the Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELISA Test kit is a combination of B. burgdorferi sensu strain B31 lysate, B. burgdorferi sensu stricto strain 2591 lysate, and a recombinant VIsE from B. burgdorferi sensu stricto strain B31. The lysates use spirochetes growing in BSK-H complete medium until mid-exponential phase. The recombinant VlsE protein is produced in E. coli SURE2 cells and purified by affinity chromatography. The purity of each antigen is assayed by SDS-PAGE followed by Coomassie staining and/or Western blotting.

## 5) Intended Use of the Device:

The Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELISA Test kit is intended as a qualitative presumptive (first step) test for the detection of IgG and IgM antibodies to B. burgdorferi sensu stricto in human serum from symptomatic patient or people suspected of infection. Positive and equivocal results must be supplemented by testing with a second-step Western blot assay.

## 6) Comparison with the Predicate Device:

The tables below provide a comparison of the Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELISA Test kit with the Trinity Biotech Captia™ Borrelia burgdorferi IgG/IgM ELISA Test kit (predicate device: K033070).

| Similarities | | |
|--------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Item | Subject Device: Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELISA Test Kit | Predicate Device: Trinity Biotech Captia™ Borrelia burgdorferi IgG/IgM ELISA Test Kit |
| Intended Use | The Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELISA Test kit is intended as a qualitative presumptive (first step) test for the detection of IgG and IgM antibodies to B. burgdorferi sensu stricto in human serum from symptomatic patients or people suspected of infection. Positive and equivocal results must be supplemented by testing with a second-step Western blot assay. | Trinity Biotech Captia™ Borrelia burgdorferi IgG/IgM ELISA Test kit is intended for the qualitative presumptive (first-step) detection of total (IgG/IgM) antibodies to B. 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 (second step) procedure. Positive supplemental (second-step) results are supportive evidence of exposure to B. burgdorferi and can be used to support a clinical diagnosis of Lyme disease. The diagnosis of Lyme disease must be made based on history, signs (such as erythema migrans), symptoms, and other laboratory data, in addition to the presence of antibodies to B. |

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| | | burgdorferi. Negative results<br>(either first or second step) should<br>not be used to exclude Lyme<br>disease. |
|-------------------|-------------------------------------------------------|------------------------------------------------------------------------------------------------------------------|
| Assay Format | Antigen coated microtiter plate – 96<br>wells. | Same |
| Technology | ELISA | Same |
| Sample Matrix | Human serum | Same |
| Controls Provided | Positive, Cutoff, Negative | Same |
| Reagents Provided | Diluent, Wash, Conjugate, Substrate,<br>Stop Solution | Same |
| Reported Results | Positive, Equivocal, Negative | Same |
| Interpretation | Optical density readings from<br>Spectrophotometer | Same |

| Differences | | |
|------------------------|------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------|
| Item | Subject Device: Gold Standard<br>Diagnostics Borrelia burgdorferi<br>IgG/IgM ELISA Test Kit | Predicate Device: Trinity Biotech<br>Captia™ Borrelia burgdorferi<br>IgG/IgM ELISA Test Kit |
| Sample Processing | Dilute Samples 1:100 in Diluent | Dilute Samples 1:20 in Diluent |
| Volumes | 100ul sample, 50ul substrate, 50ul<br>stop solution | 100ul sample, 100ul substrate,<br>100ul stop solution |
| Incubation | 15/15/15 minutes at room<br>temperature | 25/25/10-15 minutes at room<br>temperature |
| Antigens | B. burgdorferi B31 strain,<br>B. burgdorferi 2591 strain,<br>B. burgdorferi recombinant VlsE<br>B31 strain | B. burgdorferi B31 strain |
| Results Interpretation | Convert to units.<br>Negative <9<br>Equivocal 9.0-11.0<br>Positive >11.0 | Convert to units.<br>Negative ≤0.90<br>Equivocal 0.91-1.09<br>Positive ≥1.10 |

#### 6(b1): Nonclinical Studies:

#### Determination of the Assay Cutoff

The cutoff was determined by testing a total of 200 normal sera which consisted of 100 sera from an endemic region of Lyme disease and 100 sera from a non-endemic region of Lyme disease. The mean plus two standard deviations was used to determine the assay cutoff. A known positive sample was then diluted to produce a ready to use cutoff control. After the cutoff was determined 125 characterized Lyme disease samples were tested. An ROC analysis was then generated with the 325 samples (200 normal and 125 characterized samples) to verify the chosen cutoff. The analysis confirmed that the assay cutoff provided an optimal level of sensitivity and specificity.

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#### Precision

To determine the precision of the Borrelia burgdorferi IgG/IgM ELISA Test, a within-lab precision study was conducted. A precision panel consisting of a negative sample, a high negative sample, a low positive sample, and a moderate positive sample, along with the kit controls, was tested in-house. The sample panel was masked and randomized. Each of the panel members was tested in duplicate, twice per day, for 12 days. The sample panel was masked and randomized. The results are summarized in the following table:

| Sample | N | Mean<br>Units | | Within-Run | Between-Run | Between-Day | Total |
|----------------------|----|---------------|----|------------|-------------|-------------|-------|
| Moderate<br>Positive | 48 | 19.6 | SD | 0.815 | 1.534 | 1.472 | 1.737 |
| | | | CV | 4.2% | 7.8% | 7.5% | 8.9% |
| Low<br>Positive | 48 | 12.1 | SD | 0.267 | 1.417 | 1.248 | 1.442 |
| | | | CV | 2.2% | 11.7% | 10.3% | 11.9% |
| High<br>Negative | 48 | 6.1 | SD | 0.211 | 0.662 | 0.642 | 0.695 |
| | | | CV | 3.4% | 10.8% | 10.5% | 11.4% |
| Negative | 48 | 1.7 | SD | 0.113 | 0.164 | 0.151 | 0.199 |
| | | | CV | 6.6% | 9.6% | 8.8% | 11.6% |

#### Reproducibility

A reproducibility panel consisting of a negative sample, a high negative sample, a low positive sample, and a moderate positive sample, along with the kit controls, was tested at three different sites. The sample panel was masked and randomized. Each of the panel members was tested in triplicate, twice per day, for five days. The Within-Run, Between-Days, and Between-Sites Standard Deviation and Coefficients of Variation (CV) were calculated. The sample panel was masked and randomized. The results are summarized in the following table:

| Sample | N | Mean<br>Units | | Within-<br>Run | Between-<br>Run | Between-<br>Day | Between-<br>Sites | Total |
|----------------------|----|---------------|----|----------------|-----------------|-----------------|-------------------|-------|
| Moderate<br>Positive | 90 | 21.1 | SD | 1.117 | 1.543 | 1.434 | 0.386 | 1.905 |
| | | | CV | 5.3% | 7.3% | 6.8% | 1.8% | 9.0% |
| Low<br>Positive | 90 | 13.8 | SD | 0.591 | 1.155 | 1.026 | 0.404 | 1.297 |
| | | | CV | 4.3% | 8.4% | 7.5% | 2.9% | 9.4% |
| High<br>Negative | 90 | 6.4 | SD | 0.323 | 0.756 | 0.715 | 0.237 | 0.822 |
| | | | CV | 5.0% | 11.7% | 11.1% | 3.7% | 12.8% |
| Negative | 90 | 1.6 | SD | 0.145 | 0.335 | 0.312 | 0.347 | 0.365 |
| | | | CV | 9.1% | 21.1% | 19.6% | 21.8% | 23.0% |

## Analytical Specificity

The analytical specificity was determined by testing 210 asymptomatic individuals' samples from endemic (Pennsylvania) and non-endemic (Arizona) regions. The Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELISA Test results are summarized in the following table:

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| | Number of Samples | Number<br>Positive/Equivocal | Analytical Specificity |
|--------------------|-------------------|------------------------------|------------------------|
| Endemic Region | 110 | 3 | 97.3% |
| Non-endemic Region | 100 | 2 | 98.0% |

#### Cross Reactivity

A study using 221 samples was conducted to evaluate potential cross reactivity from different disease conditions. The samples were obtained from serum vendors who confirmed their positivity for each respective marker. The samples were tested on the Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELISA Test. The results are summarized in the following table:

| Infection / Diagnosis | Number of Sera<br>Tested | # Positive / (%) |
|----------------------------|--------------------------|------------------|
| Tick-borne Relapsing Fever | 26 | 2 / (7.7%) |
| Treponemal Infections | 23 | 2* (8.7%) |
| Rickettsia | 10 | 0 / (0%) |
| Ehrlichiosis | 10 | 0 / (0%) |
| Babesiosis | 11 | 0 / (0%) |
| Leptospirosis | 11 | 0 / (0%) |
| Parvovirus B19 | 12 | 0 / (0%) |
| Influenza A&B | 10 | 0 / (0%) |
| Epstein-Barr Virus | 10 | 0 / (0%) |
| Cytomegalovirus | 19 | 0 / (0%) |
| H. pylori | 11 | 0 / (0%) |
| Fibromyalgia | 10 | 1/ (10%) |
| Rheumatoid Arthritis | 11 | 0 / (0%) |
| Herpes Simplex Virus | 13 | 0 / (0%) |
| Varicella Zoster Virus | 12 | 0 / (0%) |
| Autoimmune Disease | 22 | 0 / (0%) |

*Also positive on the predicate device

#### Interfering Substances

The effect of potential interfering substances on samples using the Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELISA Test was evaluated. Three samples, a negative, a low positive and a moderate positive were spiked with high levels of interferants and were tested along with serum without spiked interferants. The recommended concentrations from the guideline "Interference Testing in Clinical Chemistry EP7-A2" from the Clinical and Laboratory Standards Institute were used (see table below). The tested substances did not affect the performance of the Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELIS A Test.

| Substance | Concentration | Interference |
|---------------|---------------|---------------|
| Albumin | 120 g/L | None detected |
| Bilirubin | 342 umol/L | None detected |
| Cholesterol | 13 mmol/L | None detected |
| Hemoglobin | 2 g/L | None detected |
| Triglycerides | 37 mmol/L | None detected |

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#### 6(b2): Clinical Studies:

#### Comparison with Predicate Device

Comparison studies were conducted at three sites (one internal and two external reference laboratories) using prospective samples submitted for Lyme serology testing. Five hundred and twenty (520) serum samples were tested on both the Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELISA Test and on the predicate B. burgdorferi IgG/IgM ELISA Test. The results are summarized in the following table:

| | | Predicate IgG/IgM ELISA | | | |
|--------------------------------------------------------------------------------|------------|-------------------------|------------|------------|------------|
| | | Positive | Equivocal* | Negative | Total |
| Gold Standard<br>Diagnostics Borrelia<br>burgdorferi IgG/IgM<br>ELISA Test Kit | Positive | 55 | 7 | 2 | <b>64</b> |
| | Equivocal* | 8 | 2 | 9 | <b>19</b> |
| | Negative | 2 | 1 | 434 | <b>437</b> |
| | Total | <b>65</b> | <b>10</b> | <b>445</b> | <b>520</b> |

*Equivocal samples counted as positive

| Positive percent agreement = 96.0% (72/75) | 95% CI (88.8% - 99.2%) |
|----------------------------------------------|------------------------|
| Negative percent agreement = 97.5% (434/445) | 95% CI (95.6% - 98.8%) |

#### Clinical Sensitivity

## Sensitivity Study

A sensitivity study was performed on 89 clinically characterized samples encompass early, disseminated, and late stages of Lyme disease. The samples were tested on both the Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELISA Test and on the predicate B. burgdorferi IgG/IgM ELISA Test. The results are summarized in the following table:

| Disease<br>Stage | n | Gold Standard<br>Diagnostics Borrelia<br>burgdorferi IgG/IgM<br>ELISA Test Kit | Predicate<br>IgG/IgM ELISA |
|------------------|----|--------------------------------------------------------------------------------|----------------------------|
| Early | 38 | 76.3% (29/38) | 78.9% (30/38) |
| Disseminated | 15 | 100.0% (15/15) | 100.0% (15/15) |
| Late | 36 | 97.2% (35/36) | 94.4% (34/36) |

## CDC Panel

Forty samples of various reactivity were acquired from the Centers for Disease Control (CDC). Of the 40 samples, five were from healthy individuals and 35 were from patients diagnose with Lyme disease and stratified by disease stage. All samples were tested on the Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELISA Test and on the predicate Borrelia burgdorferi IgG/IgM ELISA Test. The results are summarized in the following table:

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| Disease<br>Stage | n | Gold Standard Diagnostics<br><i>Borrelia burgdorferi</i><br>IgG/IgM ELISA Test Kit | | Predicate<br>IgG/IgM ELISA | |
|-------------------------------|----|------------------------------------------------------------------------------------|-------------------------------------------|-----------------------------|-------------------------------------------|
| | | Positive<br>or<br>Equivocal | % Agreement<br>with Clinical<br>Diagnosis | Positive<br>or<br>Equivocal | % Agreement<br>with Clinical<br>Diagnosis |
| Healthy | 5 | 0 | 100% | 0 | 100% |
| Early<br>(0-2 months) | 15 | 13 | 86.7% | 12 | 80.0% |
| Convalescent<br>(3-12 months) | 13 | 7 | 53.8% | 7 | 53.8% |
| Late<br>(>1 year) | 7 | 7 | 100% | 7 | 100% |

#### Expected Values

The range of values and positivity rate among different studies and population for the Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELISA Test are as follows:

| | | Unit Results | | | Qualitative Results | |
|-----------------------|-----------|--------------|-------------|--------------|-----------------------------|-----------------------------|
| Population | # Samples | Mean | Range | Std.<br>Dev. | #<br>Positive/<br>Equivocal | %<br>Positive/<br>Equivocal |
| Normal<br>Endemic | 110 | 5.8 | 0.6 – 11.3 | 2.075 | 3 | 2.7% |
| Normal<br>Non-Endemic | 100 | 4.2 | 0.9 – 12.3 | 2.077 | 2 | 2.0% |
| Prospective<br>Study | 520 | 6.3 | 0.70 - 32.3 | 5.407 | 83 | 16.0% |
| Sensitivity<br>Study | 89 | 25.0 | 3.6 - 34.9 | 8.449 | 79 | 88.8% |

Note: It is recommended that each laboratory determine its own normal range based on the population.

#### 7) Conclusion:

From the comparison data, we conclude that the Gold Standard Diagnostics Borrelia burgdorferi IgG/IgM ELISA Test is substantially equivalent to the Trinity Biotech Captia™ Borrelia burgdorferi IgG/IgM ELISA Test kit (predicate device: K033070).