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 Description

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.

AI/ML Overview

Here's a breakdown of the acceptance criteria and study details for the Gold Standard Diagnostics Borrelia burgdorferi B31 IgG Line Blot Test Kit, based on the provided 510(k) summary:

1. Table of Acceptance Criteria and Reported Device Performance

The acceptance criteria are generally implied by the predicate device comparison and various performance studies. Explicit, quantifiable acceptance criteria are not clearly stated as pass/fail thresholds in this summary but are demonstrated through observed agreement and sensitivity/specificity values compared to a predicate device and clinical classifications.

Performance Metric	Acceptance Criteria (Implied)	Reported Device Performance
Reproducibility	Consistent results for band and sample interpretation.	Band Reproducibility: Negative (100%), High Negative (92.8%), Low Positive (97.6%), Moderate Positive (100%). Sample Reproducibility: Negative (100%), High Negative (100%), Low Positive (87.8%), Moderate Positive (100%). _Note: Expected 95% for low positive_
Cross-Reactivity	Minimal or no significant cross-reactivity with common interfering conditions/infections (comparable to predicate).	3/215 (1.4%) cross-reactivity detected (2 Babesiosis, 1 Tick-borne relapsing fever/Rickettsial Diseases). These also tested positive on the predicate device.
Interfering Substances	No change in final interpretation with tested interferents.	No influence on band pattern, small variability in band intensity within normal deviation range. Final interpretation unchanged.
Specimen Stability	Consistent results after specified storage conditions.	Consistent results for: 2-8°C for up to 168 hours; Room Temp for up to 8 hours; up to 10 freeze-thaw cycles.
Clinical Sensitivity	Comparable to predicate device and clinically expected values.	Early: 7.5% (3/40) [95% CI: 1.6%-20.4%] (Predicate: 7.5%) Disseminated: 60.0% (12/20) [95% CI: 36.1%-80.9%] (Predicate: 60.0%) Late: 95.0% (38/40) [95% CI: 83.1%-99.4%] (Predicate: 92.5%)
Analytical Specificity	High specificity, especially in non-endemic regions.	Endemic: 98.3% (113/115 negative) Non-Endemic: 100% (119/119 negative)
CDC Reference Panel Agreement	High agreement with CDC reference values, comparable to predicate.	Healthy: 100% (5/5) (Predicate: 100%) Early: 93.3% (14/15) (Predicate: 100%) Intermediate: 84.6% (11/13) (Predicate: 76.9%) Late: 100% (7/7) (Predicate: 85.7%)
Method Comparison (PPA/NPA)	High Positive and Negative Percent Agreement with predicate.	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%]

2. Sample Size and Data Provenance for Test Sets

Reproducibility: Not explicitly stated as "test set" but 4 samples (negative, high negative, low positive, moderate positive) were tested, generating 30 data points per sample. Data provenance not specified (likely internal laboratory).
Cross-Reactivity: 215 specimens. Provenance: "known to contain potentially cross reactive antibodies to B. burgdorferi" and "patients with infections and sera from patients with diagnoses that can be confused with the late manifestations of Lyme disease." Retrospective implied.
Interfering Substances: 6 sera (2 positive, 1 low positive, 1 high negative, 2 negative). Data provenance not specified (likely internal laboratory).
Specimen Collection and Handling: 3 samples. Data provenance not specified (likely internal laboratory).
Sensitivity Testing: 100 clinically characterized samples. Provenance: obtained from Allen C. Steere, MD at the Massachusetts General Hospital, United States. Retrospective (clinically characterized samples).
Analytical Specificity: 234 asymptomatic samples (blood donors). Provenance: "from both endemic and non-endemic regions" (of Lyme disease), implied United States. Retrospective.
CDC Reference Panel: 40 samples (5 Healthy, 15 Early, 13 Intermediate, 7 Late). Provenance: Provided by the CDC, United States.
Method Comparison: Not explicitly stated as a single number for the "test set" but based on the 2x2 table, it's 310 samples (112+2+1+195). Provenance: Prospective clinical study at three different geographic sites in the U.S. (Pennsylvania, North Carolina, and California).

3. Number of Experts and Qualifications for Ground Truth - Test Set

Reproducibility, Cross-Reactivity, Interfering Substances, Specimen Collection & Handling: No information provided regarding experts for ground truth. Interpretation of bands for these studies was likely performed by qualified lab personnel.
Sensitivity Testing: "100 clinically characterized samples obtained from Allen C. Steere, MD at the Massachusetts General Hospital." Allen C. Steere is a highly renowned expert in Lyme disease, a physician and rheumatologist, so his clinical characterization would be considered expert ground truth. The exact number of additional experts or their qualifications is not specified, but the source implies high-level clinical expertise.
Analytical Specificity: Ground truth is implied by the samples being "asymptomatic samples (blood donors)" and their origin from endemic/non-endemic regions for Lyme disease. No explicit experts mentioned, but the nature of the samples serves as the de facto ground truth (healthy/uninfected).
CDC Reference Panel: Ground truth is established by the CDC, meaning these panels are meticulously characterized and broadly accepted as reference standards. No individual experts are named beyond the CDC institution itself.
Method Comparison: Ground truth is implied by the comparison to a "commercially available B. burgdorferi IgG Blot test" (the predicate device) after initial screening by ELISA. The study also mentions discrepant samples were tested on "a second commercially available assay." This suggests a comparative approach rather than a single expert adjudication for ground truth.

4. Adjudication Method for the Test Set

For studies where an adjudication method might be relevant (e.g., Method Comparison), the document implies a comparison to a predicate device and a secondary assay for discrepant results, rather than a formal expert adjudication method like 2+1 or 3+1. For instance, in the method comparison: "The discrepant samples were tested on a second commercially available assay." This suggests a chain of evidence but not a formal consensus.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

No, a Multi-Reader Multi-Case (MRMC) comparative effectiveness study was not explicitly done. This study focuses on the standalone performance of the device against a predicate and clinical classifications, not on how human readers' performance improves with or without AI assistance. The device is a laboratory diagnostic test, not an AI-assisted diagnostic tool for image interpretation by human readers.

6. Standalone Performance

Yes, a standalone performance study was done extensively. The entire submission details the performance of the Gold Standard Diagnostics Borrelia burgdorferi B31 IgG Line Blot Test Kit on its own, through various non-clinical and clinical tests (reproducibility, cross-reactivity, interfering substances, specimen stability, sensitivity, analytical specificity, CDC panel agreement, and method comparison with a predicate device). This is the primary focus of the 510(k) submission.

7. Type of Ground Truth Used

The ground truth varied depending on the study:

Reproducibility, Interfering Substances, Specimen Stability: Likely internal lab characterization and known properties of samples.
Cross-Reactivity: Clinical diagnosis of other infections/conditions.
Sensitivity Testing: Expert clinical characterization by Allen C. Steere, MD (Massachusetts General Hospital). This represents high-level clinical ground truth.
Analytical Specificity: Implied by the source of samples (asymptomatic blood donors from endemic/non-endemic regions), representing clinically healthy/uninfected individuals.
CDC Reference Panel: CDC-established reference values/characterization for known Lyme disease stages and healthy controls.
Method Comparison: Initial screening by ELISA, followed by comparison to a legally marketed predicate device and a "second commercially available assay" for discrepant results. This is a comparative ground truth against established methods.

8. Sample Size for the Training Set

Information on a specific "training set" is not provided in this 510(k) summary. This type of regulatory submission for a diagnostic kit typically focuses on the validation of the finalized product rather than the development and training phases of an algorithm. For a traditional laboratory assay like a line blot, "training set" doesn't apply in the same way it would for a machine learning algorithm. The development process likely involved internal optimization and testing, but these are not disclosed as formal "training sets."

9. How the Ground Truth for the Training Set Was Established

As a "training set" is not explicitly mentioned or applicable in the context of this traditional line blot assay as described in the 510(k), the method for establishing its ground truth is not applicable/not provided.

Summary

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K113847

APR 1 1 2012

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

1. 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

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).

Description of the device

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5) Intended use of the device

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. burgdorferiB31 IgG Line Blot Test Kit	Trinity Biotech B. burgdorferi (IgG)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. burgdorferiB31 IgG Line Blot Test Kit	Trinity Biotech B. burgdorferi (IgG)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	58	45	41	39	28	23	18	Numberof Bands
Negative				90			90		<4significantbands
HighNegative	90		64	90			90		4significantbands
LowPositive			90	90	90	79		90	5significantbands
ModeratePositive	90	90		90	90	90		90	>5significantbands

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 /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 ofSamples	Line Blot Sensitivitywith 95% CI	Predicate DeviceSensitivity with 95% CI
Early	40	7.5% (3/40)[1.6%-20.4%]	7.5% (3/40)[1.6%-20.4%]
Disseminated	20	60.0% (12/20)[36.1%-80.9%]	60.0% (12/20)[36.1%-80.9%]
Late	40	95.0% (38/40)[83.1%-99.4%]	92.5% (37/40)[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 ofSamples	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(0-2 months)	15	93.3% (14/15)	100% (15/15)
Intermediate(3-12 months)	13	84.6% (11/13)	76.9% (10/13)
Late(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:

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	Trinity Biotech IgG Blot
	Positive	Negative
Gold Standard DiagnosticIgG Line Blot	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 (301) 796-5450. 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 (800) 638-2041 or (301) 796-7100 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:

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)

Page 1 of

Regulation Number and Section

§ 866.3830

Treponema pallidum treponemal test reagents.(a)
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), theTreponema pallidum immobilization test (T.P.I.), and other treponemal tests used to identify antibodies toTreponema pallidum directly from infecting treponemal organisms in serum. The identification aids in the diagnosis of syphilis caused by bacteria belonging to the genusTreponema and provides epidemiological information on syphilis.(b)
Classification. Class II (performance standards).