BORRELIA BURGDORFERI IGG BLOT TEST

K113847 · Gold Standard Diagnostics · LSR · Apr 11, 2012 · Microbiology

Device Facts

• Record ID: K113847
• Device Name: BORRELIA BURGDORFERI IGG BLOT TEST
• Applicant: Gold Standard Diagnostics
• Product Code: LSR · Microbiology
• Decision Date: Apr 11, 2012
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3830
• Device Class: Class 2

Intended Use

The Gold Standard Diagnostics Borrelia burgdorferi B31 IgG Line Blot Test Kit is intended for the qualitative detection of IgG antibodies to B. burgdorferi sensu stricto (B31) in human serum. This test is intended for use in testing human serum samples which have been found positive or equivocal using an ELISA or IFA test procedure to provide supportive evidence of infection with B. burgdorferi.

Device Story

Line blot assay for qualitative detection of IgG antibodies to B. burgdorferi (B31) in human serum; utilizes purified or cloned antigenic proteins (18, 23, 28, 30, 39, 41, 45, 58, 66, 93 kDa) sprayed onto nitrocellulose strips. Patient serum incubated with strips; specific antibodies bind to antigens. Unbound components washed away; strips incubated with anti-human IgG antibody-enzyme conjugate. Substrate addition produces blue-violet precipitate at antigen-antibody complex sites. Interpretation based on observed band pattern. Used in clinical laboratory settings by trained personnel. Provides supportive evidence of Lyme disease infection following positive/equivocal ELISA or IFA screening.

Clinical Evidence

Prospective clinical study (n=310) compared subject device to predicate; positive percent agreement 99.1% (112/113), negative percent agreement 99.0% (195/197). Sensitivity study (n=100) across disease stages showed comparable performance to predicate (Early: 7.5%, Disseminated: 60.0%, Late: 95.0%). CDC reference panel testing showed high agreement across stages. Analytical specificity 98.6-100%.

Technological Characteristics

Line blot immunoassay; nitrocellulose membrane with sprayed purified/cloned B. burgdorferi proteins. Manual/semi-automated processing; enzyme-linked colorimetric detection (blue-violet precipitate). No specialized instrumentation required. Standalone diagnostic test.

Indications for Use

Indicated for qualitative detection of IgG antibodies to B. burgdorferi sensu stricto (B31) in human serum. Intended for use as a follow-up test for human serum samples previously found positive or equivocal by ELISA or IFA to provide supportive evidence of B. burgdorferi infection.

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

• B. burgdorferi (IgG) Marblot Strip Test System (K950829)

Submission Summary (Full Text)

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K113847

APR 1 1 2012

1

# 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.

| Submitter's Name: | Gold Standard Diagnostics |
|-------------------|------------------------------------|
| Address: | 2851 Spafford St. Davis, CA. 95618 |
| Phone Number: | 530-759-8000 |
| Contact Person: | Napoleon Monce |
| Date: | April 5, 2012 |

- 2) Product and Trade Name: Borrelia burgdorferi IgG B31 Line Blot Test Kit Common Name or Classification Name: Borrelia burgdorferi IgG Western blot Product Code: LSR
3) Legally marketed device to which the submitter claims equivalence

B. burgdorferi (IgG) Marblot Strip Test System (Distributed by Trinity Biotech) for the qualitative detection of human IgG antibody to individual proteins of Borrelia burgdorferi in human serum. The system is intended for use in testing human serum samples which have been found positive or equivocal using and EIA or IFA test procedure (K950829).

4) Description of the device

The assay requires a total of 70 minutes incubation time. The test uses purified antigens coated on nitrocellulose strips. Serum is added to each strip and incubated for 30 minutes. If Borrelia burgdorferi antibodies are present they will bind to the antigens on the strips. Unbound serum is removed by washing the wells three times. An enzyme-conjugated anti-human IgG is then added to each strip and incubated for 30 minutes. If antibody is present, it will bind to the antibody attached to the antigen on the strip. The wells are again washed three times followed by a DI water wash to remove any unbound conjugate. After unbound conjugate has been removed by a further washing step, a visualization of the antigen-antibody complex is accomplished by the addition of a substrate which forms blue-violet precipitates at each site (antigen bands). The reaction is stopped by washing the nitrocellulose strip with distilled or deionized water. Depending on the observed band pattern one can interpret the presence of specific IgG antibodies to B. burgdorferi infection.

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# 5) Intended use of the device

The Gold Standard Diagnostics Borrelia burgdorferi B31 IgG Line Blot Test Kit is intended for the qualitative detection of IgG antibodies to B. burgdorferi sensu stricto (B31) in human serum. This test is intended for use in testing human serum samples which have been found positive or equivocal using an ELISA or IFA test procedure to provide supportive evidence of infection with B. burgdorferi.

# 6) Comparison with the predicate device

The Gold Standard Diagnostics Borrelia Burgdorferi B31 IgG Line Blot Test Kit was compared to a commercially marketed kit by Trinity Biotech the B. burgdorferi (IgG) Marblot Strip Test System (K950829). Both kits have the same intended use and use the same methodology. Below are tables comparing the reagents and the procedures of the two devices.

## Table 1: Reagent Comparison

| Gold Standard Diagnostics B. burgdorferi<br>B31 IgG Line Blot Test Kit | Trinity Biotech B. burgdorferi (IgG)<br>Marblot Strip Test |
|------------------------------------------------------------------------|------------------------------------------------------------|
| Antigen coated nitrocellulose strips | Antigen coated nitrocellulose strips |
| Diluent/Wash Concentrate - 10x | Diluent/Wash Concentrate - 10x |
| IgG Conjugate - 100x | IgG Conjugate - 10x |
| Substrate – BCIP/NBT | Substrate |
| IgG Positive Control | IgG Band Locator Control |
| IgG Cutoff Control | IgG Weak Reactive Control |
| IgG Negative Control . | IgG Negative Control |
| Diluent/Wash Concentrate - 10x | Diluent/Wash Concentrate - 10x |

# Table 2: Procedure Comparison

| Gold Standard Diagnostics B. burgdorferi<br>B31 IgG Line Blot Test Kit | Trinity Biotech B. burgdorferi (IgG)<br>Marblot Strip Test |
|------------------------------------------------------------------------|------------------------------------------------------------|
| Dilute Samples 1:101 in Diluent/Wash | Dilute Samples 1:101 in Diluent/Wash |
| Add 15ul of Samples | Add 20ul of Samples |
| Incubate for 30 minutes while rocking | Incubate for 30 minutes while rocking |
| Wash three times with reconstituted | Wash three times with reconstituted |
| Diluent/Wash Solution | Diluent/Wash Solution |
| Add 1.5ml of reconstituted Conjugate | Add 2ml of reconstituted Conjugate |
| Incubate for 30 minutes while rocking | Incubate for 15 minutes while rocking |
| Wash three times with reconstituted | Wash three times with reconstituted |
| Diluent/Wash Solution | Diluent/Wash Solution |
| Wash one time with DI water | Wash one time with DI water |
| Add 1.5ml of Substrate | Add 2ml of Substrate |
| Incubate for 10 minutes ± 3 minutes while rocking | Incubate for 4-12 minutes while rocking |
| Wash three times with 1.5ml DI water | Wash three times with 2.0ml DI water |

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# 7) Nonclinical Tests

#### Reproducibility

The reproducibility of the assay was done by testing a negative sample, a high negative sample, a low positive sample, and a moderate positive sample in triplicate for five days, twice a day, at three sites with two technicians per site giving a total of 30 data points per sample. Results of band reproducibility and sample reproducibility are shown below:

| Sample/kDa | 93 | 66 | 58 | 45 | 41 | 39 | 30 | 28 | 23 | 18 | Number<br>of Bands |
|----------------------|----|----|----|----|----|----|----|----|----|----|----------------------------|
| Negative | | | | | 90 | | | | 90 | | <4<br>significant<br>bands |
| High<br>Negative | 90 | | | 64 | 90 | | | | 90 | | 4<br>significant<br>bands |
| Low<br>Positive | | | | 90 | 90 | 90 | | 79 | | 90 | 5<br>significant<br>bands |
| Moderate<br>Positive | 90 | | 90 | | 90 | 90 | | 90 | | 90 | >5<br>significant<br>bands |

#### Band Reproducibility:

#### Sample Reproducibility:

| Sample | Band Reproducibility | Final Interpretation | |
|-------------------|----------------------|----------------------|--------------|
| | | Positive | Negative |
| Negative | 100% (180/180) | | 100% (90/90) |
| High Negative | 92.8% (334/360) | | 100% (90/90) |
| Low Positive | 97.6% (439/450) | 87.8%* (79/90) | |
| Moderate Positive | 100% (540/540) | 100% (90/90) | |

*A low positive sample is expected to yield a positivity of 95%.

### Cross Reactivity

A cross reactivity study was performed on 215 specimens known to contain potentially cross reactive antibodies to B. burgdorferi. Sera from patients with infections and sera from patients with diagnoses that can be confused with the late manifestations of Lyme disease were tested. The results are summarized in the following table:

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| Infection / Diagnosis | Number of Sera Tested | # Positive / (%) |
|------------------------------------------------------|-----------------------|------------------|
| Tick-borne Relapsing Fever /<br>Rickettsial Diseases | 23 | 1 / (4%) |
| Treponemal Infections | 12 | 0 / (0%) |
| Ehrlichiosis | 20 | 0 / (0%) |
| Babesiosis | 20 | 2 / (10%) |
| Leptospirosis | 1 | 0 / (0%) |
| Parvovirus B19 | 9 | 0 / (0%) |
| Epstein-Barr Virus | 11 | 0 / (0%) |
| Cytomegalovirus | 32 | 0 / (0%) |
| H. pylori | 12 | 0 / (0%) |
| Fibromyalgia | 10 | 0 / (0%) |
| Rheumatoid Arthritis | 12 | 0 / (0%) |
| Herpes Simplex Virus | 16 | 0 / (0%) |
| Varicella Zoster Virus | 12 | 0 / (0%) |
| Autoimmune Disease | 25 | 0 / (0%) |

Two of the 20 Babesiosis samples and one of the 23 Tick-borne relapsing fever / Rickettsial Disease specimens were positive on the Gold Standard Diagnostics B. burgdorferi B31 IgG Line Blot Test. These samples were also tested on the predicate device which also gave positive results.

## Interfering Substances

The effect of potential interfering substances on samples using the Gold Standard Diagnostics B. burgdorferi B31 IgG Line Blot Test was evaluated. High levels of hemoglobin, bilirubin, cholesterol and intralipids in serum samples were tested on six sera (two positives, one low The recommended concentrations in the positive, one high negative, and two negatives). guideline "Interference Testing in Clinical Chemistry" from the Clinical and Laboratory Standards Institute were used. The interferents at the concentrations tested did not have any influence on the band pattern. A small variability in band intensity was seen that is in the normal

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range of deviation and did not change the final interpretation. The results are summarized in the following table:

| Substance | Concentration | Interference |
|-------------|---------------|---------------|
| Hemoglobin | 2 g/L | None detected |
| Bilirubin | 342 µmol/L | None detected |
| Cholesterol | 13 mmol/L | None detected |
| Intralipids | 37 mmol/L | None detected |

# Specimen Collection and Handling Conditions

A study was performed to substantiate the specimen storage and handling conditions for the Gold Standard Diagnostics B. burgdorferi B31 IgG Line Blot Test. Three samples were tested in triplicate after the following storage conditions:

A) 2-8ºC for 24, 48, 72, 96, 120, 144 and 168 hours.

B) Room temperature for 2, 6 and 8 hours.

C) After 1, 5 and 10 freeze and thaw cycles.

The samples gave consistent results for all testing parameters above, therefore the specimens can be held at 2-8°C for at least 168 hours or at room temperature for up to 8 hours and withstand up to 10 freeze and thaw cycles.

## 8) Clinical Testing

#### Sensitivity Testing

A sensitivity study was performed on 100 clinically characterized samples obtained from Allen C. Steere, MD at the Massachusetts General Hospital. The samples encompass early, disseminated, and late stages of Lyme disease. The results are summarized in the following table:

| | Number of<br>Samples | Line Blot Sensitivity<br>with 95% CI | Predicate Device<br>Sensitivity with 95% CI |
|--------------|----------------------|--------------------------------------|---------------------------------------------|
| Early | 40 | 7.5% (3/40)<br>[1.6%-20.4%] | 7.5% (3/40)<br>[1.6%-20.4%] |
| Disseminated | 20 | 60.0% (12/20)<br>[36.1%-80.9%] | 60.0% (12/20)<br>[36.1%-80.9%] |
| Late | 40 | 95.0% (38/40)<br>[83.1%-99.4%] | 92.5% (37/40)<br>[79.6%-98.4%] |

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# Analytical Specificity (endemic & non-endemic)

For the determination of analytical specificity, testing of 234 asymptomatic samples (blood donors) from both endemic and non-endemic regions was performed. The results are summarized in the following table:

| Region | Number of<br>Samples | Number Positive | Analytical Specificity |
|-------------|----------------------|-----------------|------------------------|
| Endemic | 115 | 2 | 98.6% |
| Non-endemic | 119 | 0 | 100% |

### CDC Reference Panel

A standard panel of positive and negative specimens provided by the Center of Disease Control (CDC) for Lyme disease detection was tested both on the Gold Standard Diagnostics B. burgdorferi B31 IgG Line Blot Test and on the predicate device. The results are summarized in the following table:

| Stage | Total | GSD Line Blot % Agreement | Predicate Device % Agreement |
|-------------------------------|-------|---------------------------|------------------------------|
| Healthy | 5 | 100% (5/5) | 100% (5/5) |
| Early<br>(0-2 months) | 15 | 93.3% (14/15) | 100% (15/15) |
| Intermediate<br>(3-12 months) | 13 | 84.6% (11/13) | 76.9% (10/13) |
| Late<br>(years) | 7 | 100% (7/7) | 85.7% (6/7) |

#### Method Comparison

The performance of the Gold Standard Diagnostics B. burgdorferi B31 IgG Line Blot test was determined by conducting a prospective clinical study at three different geographic sites (Pennsylvania, North Carolina, and California) in the U.S. The patient samples were tested by an anti-B. burgdorferi ELISA test first and the resulting equivocal and positive specimens were tested on the Gold Standard Diagnostics B. burgdorferi B31 IgG Line Blot test and a commercially available B. burgdorferi IgG Blot test. The results are summarized in the following table:

б

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| | <b>Trinity Biotech IgG Blot</b> | | |
|---------------------------------------------------|---------------------------------|----------|-----|
| | Positive | Negative | |
| <b>Gold Standard Diagnostic<br/>IgG Line Blot</b> | Positive | 112 | 2 |
| | Negative | 1 | 195 |

Positive Percent Agreement = 99.1% (112/113) [95% CI: 95.2-100%] Negative Percent Agreement = 99.0% (195/197) [95% CI: 96.4-99.9%]

The discrepant samples were tested on a second commercially available assay. On the one Line Blot negative sample that was positive by the predicate, the second assay gave a negative result. Of the two Line Blot positive and predicate negative samples, the second assay called one sample negative and the other sample positive.

## 9) Conclusion

From the performance data and kit comparison above, it is our conclusion that the Gold Standard Diagnostics B. burgdorferi B31 IgG Line Blot Test Kit is substantially equivalent to the B. burgdorferi (IgG) Marblot Strip Test System (K950829) commercially marketed by Trinity Biotech.

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# DEPARTMENT OF HEALTH & HUMAN SERVICES

Image /page/7/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES • USA" arranged around the perimeter. Inside the circle is a stylized image of three human profiles facing to the right, stacked on top of each other.

#### Food and Drug Administration

10903 New Hampshire Avenue Silver Spring, MD 20993

Gold Standard Diagnostics C/O Napoleon Monce Director, Product Development 2851 Spafford St. Davis, CA. 95618

APR 1 1 2012

Re: K113847

Trade/Device Name: Borrelia burgdorferi B31 IgG Line Blot Test Kit Regulation Number: 21 CFR 866.3830 Regulation Name: Treponema pallidum treponemal test reagents Regulatory Class: Class II Product Code: LSR Dated: December 28, 2011 Received: January 18, 2012

Dear Mr. 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.

If your device is classified (see above) into class II (Special Controls), 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 I tease be devices and I bromination that your device complies with other requirements of the Act that I DA has made a and regulations administered by other Federal agencies. You must or any I carring and the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); and good manufacturing pactice medical device-related advorse ovents) (21 cegulation (21 CFR Part 820). This letter

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## Page 2 - Napoleon Monce

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 advice for your device on our labeling regulation (21 CFR Parts 801 and 809), 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" (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 Small Manufacturers, International and Consumer Assistance at its toll-free number [phone] or [phone] or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html.

Sincerely yours,

Vägaatypa

Sally A. Hojvat, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

#### Enclosure

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# Indications for Use

- 510(k) Number (if-known): -- K1-13847 -------

Device Name: Borrelia burgdorferi B31 IgG Line Blot Test Kit

Indications For Use:

The Gold Standard Diagnostics Borrelia burgdorferi B31 IgG Line Blot Test Kit is intended for the qualitative detection of IgG antibodies to B. burgdorferi sensu stricto (B31) in human serum. This test is intended for use in testing human serum samples which have been found positive or equivocal using an ELISA or IFA test procedure to provide supportive evidence of infection with B. burgdorferi.

Prescription Use X (Part 21 CFR 801 Subpart D) AND/OR

Over-The-Counter Use (21 CFR 807 Subpart C)

(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF . NEEDED)

Concurrence of CDRH, Office of In Vitro Diagnostic Devices (OIVD)

Division Sign-Off

Office of In Vitro Diagnostic Device Evaluation and Safety

KI1384-510(k)

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