Borrelia B31 ViraChip IgG Test Kit

K163504 · Viramed Biotech AG · LSR · Jul 12, 2017 · Microbiology

Device Facts

• Record ID: K163504
• Device Name: Borrelia B31 ViraChip IgG Test Kit
• Applicant: Viramed Biotech AG
• Product Code: LSR · Microbiology
• Decision Date: Jul 12, 2017
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3830
• Device Class: Class 2

Indications for Use

The Viramed Biotech AG Borrelia B31 ViraChip® IgG Test Kit is an in vitro qualitative protein microarray assay for the detection of IgG antibodies to Borrelia burgdorferi in human serum. It is intended for use in the testing of human serum samples which have been found positive or equivocal using an EIA or IFA test procedure for B. burgdorferi antibodies. Positive results from this assay are supportive evidence of infection with B. burgdorferi, the causative agent for Lyne disease. The Borrelia B31 ViraChip® IgG Test Kit must be used with a ViraChip® Reader and the ViraChip® Software.

Device Story

The Borrelia B31 ViraChip IgG Test Kit is an in vitro protein microarray assay for detecting IgG antibodies to Borrelia burgdorferi in human serum. The device processes serum samples previously screened as positive or equivocal by EIA or IFA. The assay requires the ViraChip Reader and ViraChip Software to analyze the microarray. The system provides qualitative results to clinicians as supportive evidence for Lyme disease diagnosis. It is intended for professional use in clinical laboratory settings.

Clinical Evidence

Clinical performance evaluated via prospective study (n=128) comparing subject device to predicate immunoblot, showing 100% positive agreement and 95% negative agreement. Sensitivity study (n=98) across Lyme disease stages (EM, early neurologic, late arthritis) showed 57% sensitivity vs 60% for predicate. CDC serum panel (n=44) showed 100% agreement with reported results. Analytical specificity and cross-reactivity studies performed on 200 normal donors and 206 disease-state sera.

Technological Characteristics

Protein microarray on nitrocellulose membrane; 10 B. burgdorferi antigens immobilized as spot triplets. Enzyme immunoassay principle using alkaline-phosphatase conjugate and precipitating substrate. Requires ViraChip Reader and ViraChip Software for automated analysis. Includes internal calibrator spots and controls. Connectivity via external PC for software/calibration files.

Indications for Use

Indicated for the qualitative detection of IgG antibodies to Borrelia burgdorferi in human serum samples previously found positive or equivocal by EIA or IFA. Used as supportive evidence for Lyme disease 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.

Submission Summary (Full Text)

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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY

A. 510(k) Number: K163504

B. Purpose for Submission: To obtain a substantial equivalence determination for a new device.

C. Measurand: Anti-Borrelia burgdorferi (IgG) antibodies

D. Type of Test: Enzyme Immunoassay

E. Applicant: Viramed Biotech AG

F. Proprietary and Established Names: Borrelia B31 ViraChip IgG Test Kit

G. Regulatory Information:

1. Regulation section: 21 CFR 866.3830; Treponema pallidum treponemal test reagents

2. Classification: Class II

3. Product code: LSR; Reagent, Borrelia Serological Reagent

4. Panel: Microbiology (83)

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H. Intended Use:

1. Intended use(s): The Viramed Biotech AG Borrelia B31 ViraChip IgG Test Kit is an in vitro qualitative protein microarray assay for the detection of IgG antibodies to Borrelia burgdorferi in human serum. It is intended for use in the testing of human serum samples which have been found positive or equivocal using an EIA or IFA test procedure for B. burgdorferi antibodies. Positive results from this assay are supportive evidence of infection with B. burgdorferi, the causative agent for Lyme disease.

The Borrelia B31 ViraChip IgG Test Kit must be used with a ViraChip Reader and the ViraChip Software.

2. Indication(s) for use: Same as Intended Use

3. Special conditions for use statement(s): For prescription use only

4. Special instrument requirements: ViraChip Reader and the ViraChip Software

I. Device Description: The Viramed Biotech AG Borrelia B31 ViraChip IgG is a protein microarray assay. A protein microarray can be considered as a modified solid-phase enzyme linked immunosorbent assay. Purified B. burgdorferi antigens (93kD, 66kD, 58kD, 45kD, 41kD, 39kD, 30kD, 28kD, 23kD, and 18kD) were immobilized as individual spots onto a nitrocellulose membrane. A negative control (nc), two serum controls (sc), four conjugate controls, two for IgG and two for IgM (ccG, ccM), and six calibrator controls (cal) are also applied to each microarray. One microarray is fixed on the bottom of each cavity of a standard microtiter plate. The cavities are single breakable wells on a strip in a holding frame with 96 positions. Each Borrelia specific antigen is printed three times with the same concentration as a spot triplet. Each spot triplet corresponds to one band on an immunoblot. A schematic of the Borrelia B31 ViraChip IgG Microarray is shown below.


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For each test to be performed, the diluted test serum is added to one microarray. If specific antibodies that recognize an antigen are present, they will bind to the specific antigens on the microarray. After incubation the microarray is washed to remove unbound antibodies. Alkaline- phosphatase anti-human IgG (conjugate) is then added to each microarray and incubated. If antibodies are present, the conjugate will bind to the antibodies attached to the specific antigens. The microarray is washed to remove unbound conjugate and the substrate solution is added. If the enzyme/antibody complex is present, the substrate will undergo a precipitation and color change. After an incubation period, the reaction is stopped and the presence of precipitated substrate is visualized at specific locations on the microarray. The presence of a colored precipitation at various locations on the microarray is an indirect measurement of $B$ burgdorferi specific antibodies in the patient specimen. Visualized spots from the reaction are compared for intensity with the integrated calibrator controls for evaluation.

# J. Substantial Equivalence Information:

1. Predicate device name(s): Viramed Borrelia B31 IgG ViraStripe
2. Predicate $510(\mathrm{k})$ number(s): K092693
3. Comparison with predicate:

| Device | Predicate (K092693) |
| --- | --- |
| Similarities | |
| Reagent Comparison | |
| Nitro Cellulose Membrane Antigen Support | Nitro Cellulose Membrane Antigen Support |
| 10X Wash Sample Diluent / Wash Buffer | 10X Wash Sample Diluent / Wash Buffer |
| Chromogen/Substrate Solution RTU | Chromogen/Substrate Solution RTU |
| IgG Positive Control | IgG Positive Control |
| Negative Control | Negative Control |
| Procedure Comparison | |
| Dilute Samples 1:76 and use 100 μl dilution/well | Dilute Samples 20μl to 1.5mL |
| Dilute Controls 1:16 and use 100 μl dilution/well | Dilute Controls 100μL to 1.5mL |
| Dilute Wash Buffer 1:100 | Dilute Wash Buffer 1:100 |
| Wash with Recon buffer after Sample and Conjugate step. | Wash with Recon buffer after Sample and Conjugate step. |
| Dry Membrane/wells before reading | Dry Membrane/strips before reading |
| Differences | |
| Reading by use of the ViraChip Reader and ViraChip Software | Manual reading |

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K. Standard/Guidance Document Referenced (if applicable): N/A
L. Test Principle: Enzyme Immunoassay
M. Performance Characteristics (if/when applicable):

1. Analytical performance:

a. Precision/Reproducibility:

Precision Study: A panel of six specimens was tested by Borrelia B31 ViraChip IgG in 2 replicates, two operators per day over 12 days for a total of 48 tests for each specimen. Results were read by one ViraChip Reader. Samples were selected based on FDA cleared B. burgdorferi ELISA results, including 2 low negative samples, one high negative sample, two low positive samples and one moderate positive sample. Final positive (≥5 spots) or negative (≤4 spots) agreement was 100% for all specimens. The results of the 10 significant B. burgdorferi antigen spots are shown in the table below.

| | | | Antigens | | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Sample | Reactivity | Test results | p93 | p66 | p58 | p43 | p41 | p39 | p30 | p28 | p23 | p18 |
| Mod. Pos | Pos test results | 48 | | | | | | | | | | |
| | Neg test results | 0 | | | | | | | | | | |
| | Distinct signals | | 8 | 44 | 0 | 48 | 48 | 48 | 15 | 48 | 0 | 48 |
| | % distinct signals | | 17% | 92% | 0% | 100% | 100% | 100% | 31% | 100% | 0% | 100% |
| Low Pos | Pos test results | 0 | | | | | | | | | | |
| | Neg test results | 48 | | | | | | | | | | |
| | Distinct signals | | 0 | 0 | 0 | 0 | 48 | 0 | 0 | 0 | 48 | 0 |
| | % distinct signals | | 0% | 0% | 0% | 0% | 100% | 0% | 0% | 0% | 100% | 0% |
| Low Pos | Pos test results | 0 | | | | | | | | | | |
| | Neg test results | 48 | | | | | | | | | | |
| | Distinct signals | | 0 | 0 | 47 | 0 | 48 | 0 | 48 | 0 | 0 | 0 |
| | % distinct signals | | 0% | 0% | 98% | 0% | 100% | 0% | 100% | 0% | 0% | 0% |
| High Neg | Pos test results | 0 | | | | | | | | | | |
| | Neg test results | 48 | | | | | | | | | | |
| | Distinct signals | | 0 | 48 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| | % distinct signals | | 0% | 100% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% |
| Low Neg | Pos test results | 0 | | | | | | | | | | |
| | Neg test results | 48 | | | | | | | | | | |
| | Distinct signals | | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| | % distinct signals | | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% |
| Low Neg | Pos test results | 0 | | | | | | | | | | |
| | Neg test results | 48 | | | | | | | | | | |
| | Distinct signals | | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| | % distinct signals | | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% |

Pos = positive; Mod = moderate; Neg = negative; Distinct signals = positive spots

Reproducibility Study: A panel of six specimens was tested with the Borrelia B31 ViraChip IgG at three sites, on 5 days, by two operators, in 3 replicates, equaling a total of 90 tests per specimen. Samples were selected based on FDA cleared B.

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burgdorferi ELISA results, including 2 low negative, one high negative, two low positive, and one moderate positive sample Final positive ( $\geq 5$ spots) or negative ( $\leq 4$ spots) agreement was $100\%$ for all specimens with the exception of the low ELISA positive sample VM3076 which was found to be negative in 89 out of 90 determinations $(98.9\%)$ . The results of the 10 significant B. burgdorferi antigen spots are shown in the table below.

| | | | Antigens | | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Sample | Reactivity | Test result | p93 | p66 | p58 | p43 | p41 | p39 | p30 | p28 | p23 | p18 |
| Mod. Pos | Pos test results | 90 | | | | | | | | | | |
| | Neg test results | 0 | | | | | | | | | | |
| | Distinct signals | | 76 | 75 | 0 | 90 | 90 | 90 | 54 | 90 | 0 | 90 |
| | % distinct signals | | 84% | 83% | 0% | 100% | 100% | 100% | 60% | 100% | 0% | 100% |
| Low Pos | Pos test results | 0 | | | | | | | | | | |
| | Neg test results | 90 | | | | | | | | | | |
| | Distinct signals | | 0 | 0 | 0 | 0 | 90 | 0 | 0 | 0 | 90 | 1 |
| | % distinct signals | | 0% | 0% | 0% | 0% | 100% | 0% | 0% | 0% | 100% | 1% |
| Low Pos | Pos test results | 1 | | | | | | | | | | |
| | Neg test results | 89 | | | | | | | | | | |
| | Distinct signals | | 0 | 0 | 84 | 1 | 90 | 32 | 90 | 0 | 0 | 0 |
| | % distinct signals | | 0% | 0% | 93% | 1% | 100% | 36% | 100% | 0% | 0% | 0% |
| High Neg | Pos test results | 0 | | | | | | | | | | |
| | Neg test results | 90 | | | | | | | | | | |
| | Distinct signals | | 0 | 90 | 31 | 0 | 1 | 0 | 0 | 0 | 0 | 0 |
| | % distinct signals | | 0% | 100% | 34% | 0% | 1% | 0% | 0% | 0% | 0% | 0% |
| Low Neg | Pos test results | 0 | | | | | | | | | | |
| | Neg test results | 90 | | | | | | | | | | |
| | Distinct signals | | 0 | 0 | 0 | 0 | 1 | 1 | 0 | 0 | 0 | 0 |
| | % distinct signals | | 0% | 0% | 0% | 0% | 1% | 1% | 0% | 0% | 0% | 0% |
| Low Neg | Pos test results | 0 | | | | | | | | | | |
| | Neg test results | 90 | | | | | | | | | | |
| | Distinct signals | | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| | % distinct signals | | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% |

Pos = positive; Mod = moderate; Neg = negative; Distinct signals = positive spots

b. Linearity/assay reportable range: N/A
c. Traceability, Stability, Expected values (controls, calibrators, or methods): N/A
d. Detection limit: N/A
e. Analytical specificity:

Analytical Specificity Study: For determination of analytical specificity, 200 sera from normal blood donor individuals representing endemic and non-endemic geographic regions of the United States were tested for IgG B. burgdorferi antibodies by the Borrelia B31 ViraChip IgG.

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| | Total | Negative | Positive | % Positive | % Negative |
| --- | --- | --- | --- | --- | --- |
| Endemic | 100 | 96 | 4 | 4% | 96% |
| Non-endemic | 100 | 100 | 0 | 0% | 100% |

Cross-Reactivity Study: A total of 206 potentially cross-reactive specimens from individuals with infectious conditions or autoimmune disorders were tested on Borrelia B31 ViraChip IgG Test. The results are shown in the table below.

| Disease State Sera | Total | Borrelia B31 ViraChip IgG Positive | % Cross-reactivity |
| --- | --- | --- | --- |
| ENA autoimmune | 10 | 0 | 0% |
| Babesia microti | 10 | 3* | 30% |
| Borrelia hermsii | 6 | 1* | 17% |
| Celiac disease | 10 | 0 | 0% |
| Chlamydia trachomatis | 10 | 0 | 0% |
| Cytomegalovirus | 10 | 0 | 0% |
| Epstein–Barr virus | 10 | 0 | 0% |
| Ehrlichia chaffeensis | 10 | 2* | 20% |
| Fibromyalgia | 10 | 0 | 0% |
| Helicobacter pylori | 10 | 0 | 0% |
| Herpes simplex virus | 10 | 0 | 0% |
| Influenza | 10 | 0 | 0% |
| Leptospira interrogans | 10 | 0 | 0% |
| Lupus | 10 | 0 | 0% |
| Parvovirus B19 | 10 | 0 | 0% |
| Rheumatoid arthritis | 10 | 0 | 0% |
| Rickettsia spp. | 10 | 1* | 10% |
| Rubella virus | 10 | 0 | 0% |
| Toxoplasma gondii | 10 | 2* | 20% |
| Treponema pallidum | 10 | 0 | 0% |
| Varicella zoster virus | 10 | 0 | 0% |

* Possible co-infection with B. burgdorferi is not ruled out

Interfering Substances: Hemolyzed, lipemic, icteric, or microbially contaminated sera should not be used for testing by the Borrelia B31 ViraChip IgG Test. In addition the effect of elevated bilirubin and triglycerides on test results is not established.

f. Assay cut-off: Known positives of different levels and known negative samples were tested to determine the cut-off values for each antigen spot. Spot triplets are calculated in relation to the cut off by the ViraChip Software.

# 2. Comparison studies:

a. Method comparison with predicate device:

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Prospective Study: Three independent clinical laboratories located in Minnesota, Massachusetts, and California performed comparative testing of routinely submitted specimens for B. burgdorferi infection. The specimens testing positive or equivocal on a FDA cleared first-step EIA were tested with Borrelia B31 ViraChip IgG Test and an FDA cleared immunoblot. Interpretation of immunoblot results followed the recommended criteria described by CDC. The percent agreement with predicate device is given below.

| Borrelia B31 IgG ViraChip | Predicate Western Blot IgG | | |
| --- | --- | --- | --- |
| | Positive | Negative | Total |
| Positive | 41 | 4 | 45 |
| Negative | 0 | 83 | 83 |
| Total | 41 | 87 | 128 |
| | % Agreement | 95% Confidence Intervals |
| --- | --- | --- |
| Positive | 100% (41/41) | 91.4% - 100.0% |
| Negative | 95% (83/87) | 88.8% - 98.2% |

b. Matrix comparison: N/A

3. Clinical studies:

a. Clinical Sensitivity:

Sensitivity Study: Ninety-eight (98) sera were obtained from patients that were clinically defined and/or culture confirmed with Lyme Borreliosis; of these 98 sera, 39 were paired (20 acute and 19 convalescent) sera from patients diagnosed with erythema migrans (EM), 20 with early-disseminated Lyme disease/carditis /acute neuroborreliosis and 39 with late stage Lyme arthritis. The Borrelia B31 ViraChip IgG results are presented in comparison to the predicate device in the table below.

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| Stage of Lyme disease | Borrelia B31 ViraChip IgG | | | Predicate Western Blot IgG | |
| --- | --- | --- | --- | --- | --- |
| | Total | Positive | % Sensitivity | Positive | % Sensitivity |
| Acute EM 1-21 days from onset | 20 | 3 | 15% (3/20) | 5 | 25% (5/20) |
| Convalescent EM 4 weeks after onset | 19 | 3 | 16% (3/19) | 5 | 25%(5/20) |
| Early Neurologic | 20 | 14 | 70% (14/20) | 13 | 65% (13/20) |
| Late Arthritis | 39 | 36 | 92% (36/39) | 36 | 92% (36/39) |
| Total | 98 | 56 | 57% (56/98) | 59 | 60% (59/98) |

Sensitivity Comparison:
Borrelia B31 ViraChip IgG: 57% (56/98) (95% CI: 47.3%-66.5%)
Predicate device: 60% (59/98) (95% CI: 50.3%-69.3%)
Difference in proportion: (3/98) 3.0%

CDC Serum Panel: A Lyme disease panel containing 44 clinically defined positive and negative samples was obtained from the Centers for Disease Control and Prevention, Fort Collins, Colorado. The Borrelia B31 ViraChip IgG results for these specimens are summarized below.

| | Borrelia B31 ViraChip IgG | | | | |
| --- | --- | --- | --- | --- | --- |
| CDC Reported Results | Positive | Negative | Total | % Agreement | 95% Confidence Intervals |
| Positive | 20 | 0 | 20 | 100% (20/20) | (83.9% - 100%) |
| Negative | 0 | 24 | 24 | 100% (24/24) | (86.2% - 100%) |
| Total | 20 | 24 | 44 | - | - |

Note: The results are presented as a means to convey further information on the performance of this assay with a masked characterized serum panel from the CDC. This does not imply an endorsement of the assay by the CDC.

b. Clinical specificity: N/A
c. Other clinical supportive data (when a. and b. are not applicable): N/A

4. Clinical cut-off: N/A

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5. Expected values/Reference range:

Expected Values: The incidence of IgG antibodies to *B. burgdorferi* antigenic proteins in different patient populations tested by the Borrelia B31 ViraChip IgG Test are shown in the table below. Lyme disease specimens were obtained from patients from Wakefield/Rhode Island and Lyme/Connecticut. For the prospective studies specimens originated from areas in Massachusetts, Minnesota and California. Non-endemic blood donor samples were collected in Texas and endemic blood donor samples in Pennsylvania.

| | Antigens (% incidence) | | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Borrelia B31 ViraChip IgG spots | p93 | p66 | p58 | p45 | p41 | p39 | p30 | p28 | p23 | p18 |
| Early Lyme Disease (n=39 ) | 5% | 28% | 0% | 49% | 72% | 28% | 5% | 5% | 51% | 5% |
| Disseminated Lyme Disease | 20% | 70% | 10% | 90% | 95% | 70% | 5% | 25% | 90% | 56% |
| Late Lyme Disease (n=39 ) | 79% | 95% | 72% | 82% | 100% | 95% | 44% | 82% | 69% | 92% |
| Prospective Study (n= 128) | 24% | 62% | 3% | 38% | 70% | 50% | 3% | 20% | 44% | 35% |
| Non-Endemic Blood Donors | 6% | 32% | 0% | 2% | 42% | 27% | 1% | 0% | 10% | 4% |
| Endemic Blood Donors (n=100) | 7% | 38% | 0% | 12% | 48% | 39% | 2% | 3% | 15% | 12% |

N. Instrument Name: ViraChip Reader

O. System Descriptions:

1. Modes of Operation:

Does the applicant’s device contain the ability to transmit data to a computer, webserver, or mobile device?

☑ Yes ☐ X or No

Does the applicant’s device transmit data to a computer, webserver, or mobile device using wireless transmission?

☑ Yes ☐ No ☐ X

2. Software:

FDA has reviewed applicant’s Hazard Analysis and software development processes for this line of product types:

☑ Yes ☐ X or No

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3. Specimen Identification:

Sample IDs may be entered manually.

4. Specimen Sampling and Handling:

Samples are manually prepared according to the assay manufacturer's suggestions. The microtiter plate with lot specific factors is scanned using a 2D barcoded scanner of the packaging label. Specimens are then transferred to the assay microtiter plate for processing and analysis.

5. Calibration:

The ViraChip Reader comes with calibration files, which need to be installed on an external PC. Each microarray also contains a series of calibration spots. The measured mean intensity of the calibrator controls is multiplied by the lot specific factor for each antigen (spot triplet). The resultant value is used as cut off for the assessment of the respective antigen. A spot triplet is considered as distinct if its mean intensity is equal to or higher than the intensity of the respective cut off. A spot triplet is not assessed if its mean intensity is lower than the intensity of the respective cut off or if it is not present.

6. Quality Control:

The instrument comes with calibration settings linked to the serial number and therefore is device specific. Furthermore, every microarray has calibration spots to calculate the cut off for each antigen per well and negative/positive controls for proper assay interpretation.

P. Other Supportive Instrument Performance Characteristics Data Not Covered In The "Performance Characteristics" Section above:

Q. Proposed Labeling:

The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.

R. Conclusion:

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.