LIAISON Lyme Total Antibody Plus, LIAISON Lyme Total Antibody Plus Control Set

K193051 · DiaSorin, Inc. · LSR · Jan 29, 2020 · Microbiology

Device Facts

• Record ID: K193051
• Device Name: LIAISON Lyme Total Antibody Plus, LIAISON Lyme Total Antibody Plus Control Set
• Applicant: DiaSorin, Inc.
• Product Code: LSR · Microbiology
• Decision Date: Jan 29, 2020
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3830
• Device Class: Class 2

Intended Use

LIAISON® Lyme Total Antibody Plus: The LIAISON® Lyme Total Antibody Plus assay uses chemiluminescent immunoassay (CLIA) technology for the qualitative determination of IgG and IgM antibodies to Borrelia burgdorferi in human serum and plasma (K2-EDTA, Li-heparin) samples. This assay is intended for use on samples from patients with signs and symptoms that are consistent with Lyme disease. Positive or equivocal results should be supplemented by testing with a standardized Western blot procedure. Positive supplemental results provide evidence of exposure to B. burgdorferi and can be used to support a clinical diagnosis of Lyme disease. Negative results by LIAISON® Lyme Total Antibody Plus assay should not be used to exclude Lyme disease. The test has to be performed on the LIAISON® XL Analyzer. The LIAISON® Lyme Total Antibody Plus Control Set: The LIAISON® Lyme Total Antibody Plus Control Set is intended for use as assayed quality control samples to monitor the performance of the LIAISON® Lyme Total Antibody Plus assay. The performance characteristics of LIAISON® Lyme Total Antibody Plus Control Set have not been established for any other assays or instrument platforms different from the LIAISON® XL.

Device Story

Indirect chemiluminescent immunoassay (CLIA) for qualitative detection of IgG/IgM antibodies to Borrelia burgdorferi. Input: human serum or plasma samples. Operation: magnetic particles coated with recombinant B. burgdorferi antigens (VlsE, OspC) capture patient antibodies; mouse monoclonal anti-human IgG/IgM conjugated to isoluminol binds captured antibodies. Analyzer performs all incubation and wash steps; adds starter reagents to induce flash chemiluminescence. Output: relative light units (RLU) measured by photomultiplier, converted to index value. Used in clinical laboratories on LIAISON® XL Analyzer. Results support clinical diagnosis of Lyme disease when combined with patient history, symptoms, and supplemental Western blot testing.

Clinical Evidence

Prospective study of 1,550 clinical samples compared subject device to predicate. Negative percent agreement was 98.7% (95% CI: 97.9%-99.2%). Second-tier Western blot testing on positive/equivocal samples showed 97.9% PPA (95% CI: 89.1%-99.6%). Additional evaluation of 280 CDC-characterized samples showed high agreement across disease stages (100% for Stage III). Precision and reproducibility studies (CLSI EP05-A3/EP15-A3) demonstrated acceptable performance (total %CV < 14%). Cross-reactivity evaluated against 238 specimens from 24 disease states.

Technological Characteristics

Indirect CLIA; magnetic particle solid phase coated with recombinant B. burgdorferi antigens (VlsE, OspC); isoluminol-antibody conjugate; flash chemiluminescence detection via photomultiplier; automated on LIAISON® XL Analyzer; sample types: serum, K2-EDTA plasma, lithium heparin plasma; calibration via stored 10-point master curve with two-point verification.

Indications for Use

Indicated for qualitative detection of IgG and IgM antibodies to Borrelia burgdorferi in human serum and plasma (K₂-EDTA, Li-heparin) from symptomatic patients. Positive/equivocal results require supplemental Western blot testing. Negative results do not exclude Lyme disease.

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

• Zeus ELISA Borrelia Vlse-1/pepc10 IgG/IgM Test System (K113397)

Submission Summary (Full Text)

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Image /page/0/Picture/0 description: The image shows 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 square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

January 29, 2019

DiaSorin Inc. Mari Meyer Vice President of Regulatory & Clinical Affairs 1951 Northwestern Ave Stillwater, Minnesota 55082-0285

## Re: K193051

Trade/Device Name: LIAISON Lyme Total Antibody Plus, LIAISON Lyme Total Antibody Plus Control Set Regulation Number: 21 CFR 866.3830 Regulation Name: Treponema pallidum treponemal test reagents Regulatory Class: Class II Product Code: LSR, QCH Dated: October 31, 2019 Received: November 1, 2019

Dear Mari Meyer:

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. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. 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

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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) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; 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 https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). 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 (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email ([e-mail]) or phone [phone] or 301-796-7100).

Sincerely,

Steven Gitterman, M.D., Ph.D. Deputy Director Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

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### 5.0 510(k) SUMMARY

| SUBMITTED BY: | Mari Meyer<br>VP Regulatory and Clinical Affairs, North<br>America, DiaSorin Inc.<br>1951 Northwestern Avenue<br>Stillwater, MN 55082-0285<br>Phone [phone]<br>Fax [phone]<br>Email: [e-mail] |
|----------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| DATE PREPARED: | January 11, 2020 |
| NAME OF DEVICE: | |
| Trade Name: | LIAISON® Lyme Total Antibody Plus<br>LIAISON® Lyme Total Antibody Plus Control Set |
| Common Names/Descriptions: | Borrelia burgdorferi IgG/IgM assay and<br>Borrelia burgdorferi IgG/IgM controls |
| Classification Names: | Treponema pallidum; treponemal test reagents<br>Class II, 21 CFR: 866.3830; Microbiology |
| Product Code: | LSR |
| Predicate Device: | Zeus ELISA Borrelia VIsE1/pepC10 IgG/IgM<br>Test System (K113397) |

## INTENDED USE:

LIAISON® Lyme Total Antibody Plus: The LIAISON® Lyme Total Antibody Plus assay uses chemiluminescent immunoassay (CLIA) technology for the qualitative determination of IgG and IgM antibodies to Borrelia burgdorferi in human serum and plasma (K2-EDTA, Li-heparin) samples. This assay is intended for use on samples from patients with signs and symptoms that are consistent with Lyme disease. Positive or equivocal results should be supplemented by testing with a standardized Western blot procedure. Positive supplemental results provide evidence of exposure to B. burgdorferi and can be used to support a clinical diagnosis of Lyme disease. Negative results by LIAISON® Lyme Total Antibody Plus assay should not be used to exclude Lyme disease. The test has to be performed on the LIAISON® XL Analyzer.

The LIAISON® Lyme Total Antibody Plus Control Set: The LIAISON® Lyme Total Antibody Plus Control Set is intended for use as assayed quality control samples to monitor the performance of the LIAISON® Lyme Total Antibody Plus assay. The performance characteristics of LIAISON® Lyme Total Antibody Plus Control Set have not been established for any other assays or instrument platforms different from the LIAISON® XL.

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## KIT DESCRIPTION:

The method for qualitative determination of IgG and IgM antibodies to Borrelia burgdorferi is an indirect chemiluminescence immunoassay (CLIA). All assay steps (with the exception of magnetic particle resuspension) and incubations are performed by the Analyzer. The principal components of the test are magnetic particles (solid phase) coated with recombinant Borrelia antigens and a conjugate reagent containing two mouse monoclonal antibodies (anti- human IgG and anti-human IgM) linked to an isoluminol derivative (isoluminol-antibody conjugate). During the first incubation, antigen-specific antibodies present in calibrators, samples or controls bind to the solid phase. During the second incubation, the conjuqates react with Borrelia burgdorferi IgG and IgM antibodies captured by the solid phase. Unbound material is removed with a wash cycle following incubations. Subsequently, the starter reagents are added and a flash chemiluminescence reaction is thus induced. The light signal, and hence the amount of isoluminol-antibody conjugate, is measured by a photomultiplier as relative light units (RLU) and is indicative of the presence of Borrelia burgdorferi antibodies present in calibrators, samples or controls.

| Table 1: Table of Similarities | | |
|--------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Characteristic | Candidate Device LIAISON®<br>Lyme Total Antibody Plus | Predicate Device<br>Zeus ELISA Borrelia VIsE1/pepC10<br>IgG/IgM Test System - K113397 |
| Intended Use | LIAISON® Lyme Total Antibody Plus:<br>The LIAISON® Lyme Total Antibody<br>Plus assay uses chemiluminescent<br>immunoassay (CLIA) technology for<br>the qualitative determination of IgG<br>and IgM antibodies of Borrelia<br>burgdorferi in human serum and<br>plasma (K2-EDTA, Li-heparin)<br>samples. This assay is intended for<br>use on samples from patients with<br>signs and symptoms that are<br>consistent with Lyme disease.<br>Positive or equivocal results should<br>be supplemented by testing with a<br>standardized Western blot procedure.<br>Positive supplemental results provide<br>evidence of exposure to B.<br>burgdorferi and can be used to<br>support a clinical diagnosis of Lyme<br>disease. Negative results by<br>LIAISON® Lyme Total Antibody Plus<br>assay should not be used to exclude<br>Lyme disease. The test has to be<br>performed on the LIAISON® XL<br>Analyzer. | Qualitative detection of IgG and IgM class<br>antibodies to VIsE1 and pepC10 antigens<br>from Borrelia burgdorferi in human serum.<br>The assay is intended for testing serum<br>samples from symptomatic patients or<br>those with a history of Lyme Borreliosis. All<br>positive and equivocal specimens should<br>be tested with a second-tier test such as<br>Western Blot, which if positive, is supportive<br>evidence of infection with Borrelia<br>burgdorferi. Diagnosis of Lyme<br>Borreliosis should be made based on the<br>presence of B. burgdorferi antibodies,<br>history, symptoms, and other<br>laboratory data. Negative first or second<br>tier results should not be used to exclude<br>Borreliosis. This kit is for in vitro diagnostic<br>use. |
| Results | Qualitative | Same |

# COMPARISON TO PREDICATE DEVICE

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| Measurand | IgG and IgM antibodies to Borrelia<br>burgdorferi | Same |
|-------------------------------------|----------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------|
| Intended Population | Patients with signs and symptoms<br>consistent with Borrelia infection<br>(Lyme disease) | Same |
| Assay Principle | Uses Borrelia antigens coated on a<br>solid phase to capture specific patient<br>antibodies. | Uses B. burgdorferi antigen coated on a<br>solid phase to capture specific patient<br>antibodies. |
| Sample Type | Human serum, serum separator<br>tubes, K₂-EDTA, lithium heparin plasma | Human serum |
| Conjugate antibody<br>specificities | Anti-human IgG and anti-human IgM | Anti-human IgG/IgM |
| Assay Output | Index | Same |

| Table 2: | Table of Differences | |
|--------------------|------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------|
| Feature | Candidate Device<br>LIAISON® Lyme Total<br>Antibody Plus | Predicate Device<br>Zeus ELISA Borrelia<br>VlsE1/pepC10 IgG/IgM Test |
| Test Format | CLIA (indirect chemiluminescent<br>assay) | ELISA |
| Reporter Molecule | Isoluminol derivative conjugated to<br>anti-human IgG and IgM | TMB (as a substrate for Horseradish<br>peroxidase conjugated to anti-human<br>IgG/IgM). |
| Antigen | Recombinant VlsE antigen from B.<br>burgdorferi strain B31 and from B.<br>garinii strain Pbi, and OspC antigen<br>from Borrelia afzelii, | VlsE1 and pepC10 antigens of B.<br>burgdorferi |
| Assay Procedure | Automated (on the LIAISON® XL<br>Analyzer) | Manual |
| Calibration | Two-point verification (in triplicate) of<br>stored 10-point master curve | Single Cut-off Calibrator assayed in<br>triplicate |
| Output Signal | Flash chemiluminescent response is<br>integrated over a 3 second reading<br>period to generate a relative light<br>unit (RLU) value. | Microtiter well O.D. (450 nm) is measured<br>after the enzyme reaction is halted by<br>sulfuric acid. |
| Measurement System | Photomultiplier (flash<br>chemiluminescence reader) | Spectrophotometer (EIA Microtiter plate<br>reader) |

## PERFORMANCE DATA:

Prospective Study/Method Comparison Agreement:

One thousand five hundred fifty (1550) samples collected from subjects sent to the lab for Lyme disease testing, were de-identified and numbered. The collection consisted of subjects from five (5) geographical regions of the U.S. The samples were tested with the LIAISON® Lyme Total Antibody Plus assay on the LIASON® XL and performed in three (3) laboratories (2 external and internally at DiaSorin). Results were evaluated for first tier testing.

## Table 3: First Tier Percent Agreement with Predicate Device

Predicate Assay (IgG/IgM)

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| LIAISON®<br>Lyme Total<br>Antibody Plus | Positive | Equivocal | Negative | Total |
|-----------------------------------------|----------|-----------|----------|-------|
| Positive | 62 | 1 | 9 | 72 |
| Equivocal | 2 | 0 | 10 | 12 |
| Negative | 42 | 8 | 1416 | 1466 |
| Total | 106 | 9 | 1435 | 1550 |

Positive % Agreement* 56.5% (65/115) 95% Cl: 47.4% - 65.2% Negative % Agreement 98.7% (1416/1435) 95% Cl: 97.9% - 99.2% *Includes Positive and Equivocal combined

# Table 4: Second Tier Testing

Western blot testing was performed on the samples positive or equivocal by the test device and the predicate. The following results were obtained:

| Test System | Tier 1 + or Eqv | Western Blot<br>lgG/lgM + | Western<br>Blot<br>lgG/IgM - |
|-----------------------------------------------|-----------------|---------------------------|------------------------------|
| LIAISON® Lyme Total Antibody Plus | 84 | 48 | 36 |
| Predicate Assay | 115 | 48 | 67 |
| Predicate + LIAISON® Lyme Total Antibody Plus | 65 | 47 | 18 |

Agreement Results:

2nd Tier PPA 97.9% (47/48) 95% Cl: 89.1%-99.6%

## 15.2 Characterized Lyme Panel

Two hundred eighty samples of various reactivity were acquired from the CDC and evaluated internally at the manufacturer's site. The results of the testing are presented here as a means of conveying further information on the performance of the LIAISON® Lyme Total Ab Plus assay with a characterized serum panel. This does not imply an endorsement of the assay by the CDC.

## Table 5: Testing of CDC Lyme Reference Sera

| Sample Category (CDC<br>Reference Classification) | Candidate | | | Predicate | | | N | LIAISON Lyme<br>Total AB Plus<br>% Agreement/<br>95% Wilson CI | Predicate<br>% Agreement/<br>95% Wilson CI |
|---------------------------------------------------|-----------|-----|-----|-----------|-----|-----|-----|----------------------------------------------------------------|--------------------------------------------|
| Stage I<br>Acute | Pos | Neg | Eqv | Pos | Neg | Eqv | | | |
| Stage I Acute | 27 | 9 | 3 | 30 | 9 | 0 | 39 | 76.9% (30/39)<br>61.7% - 87.4% | 76.9% (30/39)<br>61.7% - 87.4% |
| Stage II Convalescent | 29 | 2 | 0 | 29 | 2 | 0 | 31 | 93.5% (29/31)<br>79.3% - 98.2% | 93.5% (29/31)<br>79.3% - 98.2% |
| Stage III Late | 20 | 0 | 0 | 20 | 0 | 0 | 20 | 100.0% (20/20)<br>83.9% - 100.0% | 100.0% (20/20)<br>83.9% - 100.0% |
| Look-alike Diseases | 2 | 86 | 2 | 4 | 85 | 1 | 90 | 95.6% (86/90)<br>89.1% - 98.3% | 94.4% (85/90)<br>87.6% - 97.6% |
| Healthy Controls | 2 | 98 | 0 | 3 | 95 | 2 | 100 | 98.0% (98/100)<br>93.0% - 99.4% | 95.0% (95/100)<br>88.8% - 97.8% |

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### 15.3 PRECISION STUDY

A 12-day precision/repeatability study was conducted at DiaSorin on the LIAISON® Lyme Total Antibody Plus assay. Six (6) serum samples and one (1) lot of LIAISON® Lyme Total Antibody Plus Controls were tested for 12 days, 2 runs/day, and 2 replicates per run by multiple technologists for a total of 48 replicates. These test days span 2 calibration cycles. CLSI document EP05-A3 was consulted in the preparation of the testing protocol.

| Sample ID | N | Mean<br>(Index) | Within Run | | Between Run | | Between Day | | TOTAL | |
|----------------|----|-----------------|------------|-----|-------------|-----|-------------|-----|-------|------|
| | | | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Neg<br>Control | 48 | 0.09 | 0.01 | 9.4 | 0.01 | 8.0 | 0.00 | 0.0 | 0.01 | 11.3 |
| Pos Control | 48 | 1.96 | 0.13 | 6.7 | 0.03 | 1.6 | 0.15 | 7.7 | 0.20 | 10.4 |
| Sample 1 | 48 | 0.09 | 0.00 | 5.6 | 0.01 | 6.0 | 0.00 | 2.2 | 0.01 | 8.5 |
| Sample 2 | 48 | 0.84 | 0.04 | 4.5 | 0.02 | 2.8 | 0.05 | 5.6 | 0.07 | 7.7 |
| Sample 3 | 48 | 1.59 | 0.12 | 7.7 | 0.00 | 0.0 | 0.10 | 6.0 | 0.15 | 9.4 |
| Sample 4 | 48 | 4.51 | 0.21 | 4.7 | 0.24 | 5.4 | 0.25 | 5.5 | 0.41 | 9.0 |
| Sample 5 | 48 | 0.82 | 0.06 | 7.3 | 0.07 | 8.4 | 0.06 | 7.0 | 0.11 | 13.2 |
| Sample 6 | 48 | 1.39 | 0.08 | 6.0 | 0.12 | 8.7 | 0.07 | 5.2 | 0.16 | 11.8 |

### 15.4 REPRODUCIBILITY STUDY

A five (5) day precision/reproducibility study was performed internally at DiaSorin Inc. and at two (2) external U.S. laboratories with one (1) lot of LIAISON® Lyme Total Antibody Plus assay. The study was performed for 5 days, 2 runs/day, and 3 replicates/run. Each day, two operators, at each testing site performed the testing for a total of 30 replicates at each site. CLSI document EP15-A3 was consulted in the preparation of the testing protocol.

| Sample | | | Within Run | | Between Day | | Between Run | | Between Site | | TOTAL | |
|-------------|----|--------|------------|-----|-------------|-----|-------------|-----|--------------|-----|-------|-----|
| ID | n | mean | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Neg Control | 90 | 0.0509 | 0.002 | 4.7 | 0.002 | 3.4 | 0.002 | 3.4 | 0.002 | 4.7 | 0.004 | 8.2 |
| Pos Control | 90 | 1.8 | 0.118 | 6.5 | 0.04 | 2.2 | 0.039 | 2.2 | 0.000 | 0.0 | 0.13 | 7.2 |
| Sample 1 | 90 | 0.0463 | 0.002 | 4.6 | 0.002 | 4.3 | 0.001 | 3.1 | 0.000 | 0.0 | 0.003 | 7.0 |
| Sample 2 | 90 | 0.654 | 0.028 | 4.2 | 0.036 | 5.5 | 0.026 | 3.9 | 0.024 | 3.7 | 0.057 | 8.8 |
| Sample 3 | 90 | 1.55 | 0.07 | 4.5 | 0.055 | 3.6 | 0.046 | 3.0 | 0.077 | 5.0 | 0.126 | 8.2 |
| Sample 4 | 90 | 4.66 | 0.207 | 4.4 | 0.161 | 3.5 | 0.107 | 2.3 | 0.000 | 0.0 | 0.283 | 6.1 |
| Sample 5 | 90 | 0.86 | 0.052 | 6.1 | 0.04 | 4.6 | 0.000 | 0.0 | 0.035 | 4.1 | 0.074 | 8.7 |
| Sample 6 | 90 | 1.42 | 0.065 | 4.6 | 0.075 | 5.3 | 0.041 | 2.9 | 0.064 | 4.5 | 0.125 | 8.8 |

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### 15.5 Cross-Reactivity Study

The cross-reactivity study was designed to evaluate 238 specimens from twenty four (24) disease states either known to contain potentially cross-reactive antibodies to B. burgdorferi or from patients with diagnoses that can exhibit signs and symptoms similar to late manifestations of Lyme disease and cause false positive results.

| Organism Infected or Disease State | Samples<br>Tested<br>(n) | Pos or Eqv |
|------------------------------------------------------------|--------------------------|------------|
| Tick Borne Diseases | | |
| Babesiosis | 10 | 4 |
| Tick Borne Relapsing Fever (TBRF) | 8 | 2 |
| Autoimmune Disorders | | |
| Anti-Nuclear Antibodies (ANA) | 10 | 0 |
| Multiple Sclerosis | 10 | 0 |
| Sjogrens Syndrome | 10 | 1 |
| Viral Diseases | | |
| Cytomegalovirus (CMV) IgM | 10 | 0 |
| Cytomegalovirus (CMV) IgG | 10 | 0 |
| Epstein-Barr Virus (EBV) VCA, and/or<br>heterophile Ab IgM | 10 | 0 |
| Epstein-Barr Virus (EBV) VCA, NA-1 and/or<br>EA-D IgG | 10 | 0 |
| Epstein-Barr Virus (EBV) EBNA IgG | 10 | 1 |
| Epstein-Barr Virus (EBV) VCA IgM | 10 | 2 |
| Epstein-Barr Virus (EBV) VCA IgG | 10 | 0 |
| Human Immunodeficiency Virus (HIV) | 10 | 0 |
| Influenza Virus | 10 | 0 |
| Parvovirus | 10 | 3 |
| Bacterial Diseases | | |
| E. coli | 10 | 0 |
| H. pylori | 10 | 0 |
| Syphilis | 10 | 0 |
| Rheumatic Diseases | | |
| Fibromyalgia | 10 | 0 |
| Rheumatoid Arthritis | 10 | 0 |
| Rheumatoid Factor | 10 | 0 |
| Systemic Lupus Erythematosus (SLE) | 10 | 0 |
| Additional Markers | | |
| Chronic Fatigue Syndrome | 10 | 0 |
| Human Anti-mouse Antibodies (HAMA) | 10 | 0 |
| Total | 238 | 13 |

### 15.6 Interfering Substances

Controlled studies of potentially interfering substances from endogenous interferents spiked into equivocal B. burgdorferi serum specimens showed that assay performance was not affected at the concentration for each substance listed below. The testing was based on CLSI-EP7-A3.

| Substance | Concentration |
|---------------|---------------|
| Hemoglobin | 1000 mg/dL |
| Triglycerides | 1500 mg/dL |
| Bilirubin | 40 mg/dL |
| Total protein | 12 g/dL |
| Cholesterol | 500 mg/dL |
| Biotin | 3600 ng/mL |

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#### 15.7 Matrix Equivalence Study

Thirty-two (32) matched patient sets of serum, SST serum, K2-EDTA plasma and lithium heparin plasma samples were tested to determine if these sample types provide equivalent results. Sample regression analysis was done by Passing and Bablok method. All sample types met acceptance criteria for use in the LIAISON® Lyme Total Antibody Plus assay. A summary of the results is shown in the following table.

Sample Equivalence Results:

| Comparison to Serum | Bias | CI: 95% | |
|------------------------------|------|---------|------|
| SST Serum Constant | 0.00 | 0.01 | 0.01 |
| SST Serum Proportional | 0.99 | 0.97 | 1.01 |
| K2-EDTA Constant | 0.01 | 0.03 | 0.01 |
| K2-EDTA Proportional | 0.99 | 0.95 | 1.01 |
| Lithium Heparin Constant | 0.01 | 0.02 | 0.01 |
| Lithium Heparin Proportional | 0.95 | 0.92 | 0.97 |

## CONCLUSION:

The material submitted in this premarket notification supports a substantial equivalence decision.