Viramed Borrela All-In-One ViraChip Test Kit

{0} FDA U.S. FOOD &amp; DRUG ADMINISTRATION # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY ## I Background Information: A 510(k) Number K220016 B Applicant Viramed Biotech AG C Proprietary and Established Names Viramed Borrelia All-In-One ViraChip Test Kit D Regulatory Information | Product Code(s) | Classification | Regulation Section | Panel | | --- | --- | --- | --- | | LSR | Class II | 21 CFR 866.3830 - Treponema Pallidum Treponemal Test Reagents | MI - Microbiology | ## II Submission/Device Overview: A Purpose for Submission: To obtain a substantial equivalence determination for a new device. B Measurand: Anti-Borrelia burgdorferi (IgG and IgM) antibodies C Type of Test: Enzyme Immunoassay Food and Drug Administration 10903 New Hampshire Avenue Silver Spring, MD 20993-0002 www.fda.gov {1} III Intended Use/Indications for Use: A Intended Use(s): See Indications for Use below. B Indication(s) for Use: The Viramed Biotech AG Borrelia All-In-One ViraChip is an in vitro qualitative microarray assay for the detection of IgM and IgG antibodies to *Borrelia burgdorferi* in human serum. The assay is intended for testing serum samples from symptomatic patients or those suspected of Lyme Disease. It is intended to detect antibodies to VlsE and multiple other *B. burgdorferi* antigens following a modified two-tier test methodology. Positive results from the Viramed Biotech AG Borrelia All-In-One ViraChip are supportive evidence for the presence of antibodies and exposure to *B. burgdorferi*, the causative agent for Lyme disease. Negative results do not preclude infection with *B. burgdorferi*. Test results are to be used in conjunction with information obtained from the patient’s clinical evaluation and other diagnostic procedures as an aid in diagnosis of Lyme disease. The Viramed Biotech AG Borrelia All-In-One ViraChip Test must be used with a ViraChip Reader and the ViraChip Software. C Special Conditions for Use Statement(s): Rx - For Prescription Use Only D Special Instrument Requirements: ViraChip Reader and the ViraChip Software IV Device/System Characteristics: A Device Description: The Viramed Biotech AG Borrelia All-In-One ViraChip Test Kit is a protein microarray assay. A protein microarray can be considered as a modified solid-phase enzyme linked immunosorbent assay. Isolated *B. burgdorferi* antigens are bound to a solid phase nitrocellulose support membrane. Purified *B. burgdorferi* VlsE and *B. burgdorferi* antigens with the following molecular weights are used: 93 kDa, 58 kDa, 45 kDa, 39 kDa, 30 kDa, 23 kDa, 21 kDa, 19 kDa, 18 kDa and 17 kDa. The antigens were immobilized as individual spot triplets onto the nitrocellulose membrane. Positions of the spots are exactly defined and can be assigned to the antigen reliably. A negative control (nc), two serum controls (sc), four conjugate controls (ccG and 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 in a holding frame with 96 positions. A schematic of the Borrelia All-In-One ViraChip is shown below. K220016 - Page 2 of 14 {2}  Figure 1. Borrellia All-In-One ViraChip Microarray # B Principle of Operation: For each test to be performed, the diluted test serum sample 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-conjugated anti-human antibodies (conjugate) are then added to each microarray and incubated. If host 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 Borrelia burgdorferi specific antibodies in the patient specimen. Visualized spots from the reaction are compared for intensity with the integrated calibrator controls for evaluation. Spot intensity measurements are performed by the ViraChip Reader and that the assay result is interpreted by the ViraChip Software. The assay result interpretation algorithm is described in the table below. Test results are considered positive if the VlsE spot triplet is positive or equivocal and at least one additional spot triplet is positive. For all test results, the VlsE spots and intensities are shown in the ViraChip Software on the screen, in the print-outs from the ViraChip Software, and in the exported files. All other antigen spots and their corresponding intensities are only reported if the VlsE spot triplet is positive or equivocal. If the VlsE spot triplet is negative, all other antigen spots are masked by white circles. K220016 - Page 3 of 14 {3} Table 1. Borrelia All-In-One ViraChip Interpretation of Results | VlsE Spot Triplet | | 93 kDa, 58 kDa, 45 kDa, 39 kDa, 30 kDa, 23 kDa, 21 kDa, 19 kDa, 18 kDa and 17 kDa Spot Triplets | | Borrelia All-In-One ViraChip Result | | --- | --- | --- | --- | --- | | Spot Intensities | Spot Triplet Result | Spot Intensities | Spot Triplet Result | | | ≥ 100 units | Positive | ≥ 100 units for at least one spot triplet out of 93, 58, 45, 39, 30, 23, 21, 19, or 17kDa Or ≥ 90 units for spot triplet 18 kDa | Positive | Positive | | | | < 100 units for all spot triplets AND < 90 units for spot triplet 18 kDa | Negative | Negative | | ≥ 60 and < 100 units | Equivocal | ≥ 100 units for at least one spot triplet out of 93, 58, 45, 39, 30, 23, 21, 19, or 17kDa Or ≥ 90 units for spot triplet 18 kDa | Positive | Positive | | | | < 100 units for all spot triplets AND < 90 units for spot triplet 18 kDa | Negative | Negative | | < 60 units | Negative | Not interpreted | Not interpreted | Negative | K220016 - Page 4 of 14 {4} V Substantial Equivalence Information: A Predicate Device Name(s): Borrelia B31 ViraChip IgG Test Kit, Borrelia B31 ViraChip IgM Test Kit, ZEUS ELISA Borrelia VlsE1/PepC10 IgG/IgM Test System B Predicate 510(k) Number(s): K163504, K163695, K113397 C Comparison with Predicate(s): | Device & Predicate Device(s): | K220016 | K163504 | K163695 | | --- | --- | --- | --- | | Device Trade Name | Viramed Borrelia All-In-One ViraChip Test Kit | Borrelia B31 ViraChip IgG Test Kit | Borrelia B31 ViraChip IgM Test Kit | | General Device Characteristic Similarities | | | | | Intended Use/Indications For Use | The Viramed Biotech AG Borrelia All-In-One ViraChip is an in vitro qualitative microarray assay for the detection of IgM and IgG antibodies to Borrelia burgdorferi in human serum. The assay is intended for testing serum samples from symptomatic patients or those suspected of Lyme Disease. It is intended to detect antibodies to VlsE and multiple other B. burgdorferi antigens following a modified two-tier test methodology. Positive results from the Viramed Biotech AG Borrelia All-In-One ViraChip are supportive evidence for the presence of antibodies and exposure to B. burgdorferi, the causative agent for | 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. | The Viramed Biotech AG Borrelia B31 ViraChip IgM Test Kit is an in vitro qualitative protein microarray assay for the detection of IgM 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 Viramed Biotech AG Borrelia B31 ViraChip IgM Test Kit must be used with a ViraChip Reader and the ViraChip Software. | K220016 - Page 5 of 14 {5} K220016 - Page 6 of 14 | | Lyme disease. Negative results do not preclude infection with B. burgdorferi. Test results are to be used in conjunction with information obtained from the patient’s clinical evaluation and other diagnostic procedures as an aid in diagnosis of Lyme disease. The Viramed Biotech AG Borrelia All-In-One ViraChip Test must be used with a ViraChip Reader and the ViraChip Software. | | | | --- | --- | --- | --- | | Assay Design | Antigen Coated wells (Microarrays) | Same | Same | | Reagents | 10X Wash Buffer, Sample Buffer, Sample Buffer, Chromogen/Substrate Solution | Same | Same | | Sample Volume | Samples diluted 1:76 and 100 µL added per well | Same | Same | | Procedural Steps | Wash after Sample and Conjugate Step | Same | Same | | Controls | Positive Control Serum, Negative Control Serum | Same | Same | | Result Generation | Automated with ViraChip Reader | Same | Same | | **General Device Characteristic Differences** | | | | | Antigens | VlsE, 93 kD, 58 kD, 45 kD, 39 kD, 30 kD, 23 kD, 21 kD, 19 kD, 18 kD, and 17 kD | 93 kD, 66 kD. 58 kD, 45 kD, 41 kD, 39 kD, 30 kD, 28 kD, 23 kD, and 18 kD | 41 kD, 39 kD, and 23 kD | | Antibodies Detected | IgG and IgM | IgG | IgM | {6} K220016 - Page 7 of 14 | Device & Predicate Device(s): | K220016 | K113397 | | --- | --- | --- | | Device Trade Name | Viramed Borrelia All-In-One ViraChip Test Kit | ZEUS ELISA Borrelia VlsE1/PepC10 IgG/IgM Test System | | General Device Characteristic Similarities | | | | Intended Use/Indications For Use | The Viramed Biotech AG Borrelia All-In-One ViraChip is an in vitro qualitative microarray assay for the detection of IgM and IgG antibodies to *Borrelia burgdorferi* in human serum. The assay is intended for testing serum samples from symptomatic patients or those suspected of Lyme Disease. It is intended to detect antibodies to VlsE and multiple other *B. burgdorferi* antigens following a modified two-tier test methodology. Positive results from the Viramed Biotech AG Borrelia All-In-One ViraChip are supportive evidence for the presence of antibodies and exposure to *B. burgdorferi*, the causative agent for Lyme disease. Negative results do not preclude infection with *B. burgdorferi*. Test results are to be used in conjunction with information obtained from the patient’s clinical evaluation and other diagnostic | The ZEUS ELISA Borrelia VlsE1/PepC10 IgG/IgM Test System is intended for the qualitative detection of IgG and IgM class antibodies to VlsE1 and pepC10 antigens from *Borrelia burgdorferi* in human serum. The assay is intended for testing serum samples from symptomatic patients or those with a history of Lyme Borreliosis. All positive and equivocal specimens should be tested with a second-tier test such as Western Blot, which if positive, is supportive evidence of infection with *Borrelia burgdorferi*. Diagnosis of Lyme Borreliosis should be made based on the presence of *B. burgdorferi* antibodies, history, symptoms, and other laboratory data. Negative first or second tier results should not be used to exclude Borreliosis. This kit is for in vitro diagnostic use. | {7} | | procedures as an aid in diagnosis of Lyme disease. The Viramed Biotech AG Borrelia All-In-One ViraChip Test must be used with a ViraChip Reader and the ViraChip Software. | | | --- | --- | --- | | Antibodies Detected | IgG and IgM | Same | | Controls | Positive Control Serum, Negative Control Serum | Same | | General Device Characteristic Differences | | | | Antigens | VlsE, 93 kD, 58 kD, 45 kD, 39 kD, 30 kD, 23 kD, 21 kD, 19 kD, 18 kD, and 17 kD | Recombinant VlsE antigen and synthetic pepC10 antigen | | Assay Design | Antigen Coated wells (Microarrays) | ELISA | | Procedural Steps | Wash after Sample and Conjugate Step | Wash between sample and conjugate incubation steps, incubate with substrate | | Reagents | 10X Wash Buffer, Sample Buffer, Sample Buffer, Chromogen/Substrate Solution | Calibrator, SAVe Diluent, TMB substrate solution, Stop Solution, Wash buffer concentrate | | Result Generation | Automated with ViraChip Reader | ELISA microwell reager, 450 nm | | Sample Volume | Samples diluted 1:76 and 100 μL added per well | Sample diluted 1:21 in SAVe Diluent | VI Standards/Guidance Documents Referenced: M34-1A: Western Blot Assay for Antibodies to Borrelia Burgdorferi; Approved Guideline EP05-A3 Evaluation of Precision Performance of Quantitative Measurement Methods; Approved Guideline- Third Edition VII Performance Characteristics (if/when applicable): K220016 - Page 8 of 14 {8} # A Analytical Performance: # 1. Precision/Reproducibility: # Precision Study A panel of eight specimens was tested by the Borrelia All-In-One ViraChip in two replicates by two operators per day over 12 days for a total of 48 tests for each specimen. Samples were selected based on FDA cleared B. burgdorferi antibody ELISA results, including one low negative, one moderate negative, one high negative one low positive, two moderate positive, and one high positive sample. The relative intensity of each antigen was classified as distinct signal with the thresholds as described in the interpretation criteria. Final positive and negative agreement was $100\%$ for all specimens with the exception of a high negative specimen which exhibited a positive result for 6/48 replicates. Table 2. Precision Study Results Summary | | | | Antigens | | | | | | | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | Sample | Reactivity | Test results | VlsE | p93 | p58 | p45 | p39 | p30 | p23 | p21 | p19 | p18 | p17 | | High Pos | Poss test results | 48 | | | | | | | | | | | | | | Neg test results | 0 | | | | | | | | | | | | | | Distinct signals | | 48 | 0 | 45 | 0 | 48 | 48 | 48 | 48 | 10 | 48 | 0 | | | % distinct signals | | 100% | 0% | 94% | 0% | 100% | 100% | 100% | 100% | 21% | 100% | 0% | | Mod. Pos | Pos test results | 48 | | | | | | | | | | | | | | Neg test results | 0 | | | | | | | | | | | | | | Distinct signals | | 48 | 0 | 0 | 48 | 0 | 48 | 0 | 45 | 48 | 48 | 48 | | | % distinct signals | | 100% | 0% | 0% | 100% | 0% | 100% | 0% | 94% | 100% | 100% | 100% | | Mod Pos | Pos test results | 48 | | | | | | | | | | | | | | Neg test results | 0 | | | | | | | | | | | | | | Distinct signals | | 48 | 48 | 48 | 0 | 48 | 0 | 0 | 48 | 0 | 48 | 0 | | | % distinct signals | | 100% | 100% | 100% | 0% | 100% | 0% | 0% | 100% | 0% | 100% | 0% | | Mod Pos | Pos test results | 48 | | | | | | | | | | | | | | Neg test results | 0 | | | | | | | | | | | | | | Distinct signals | | 48 | 0 | 0 | 0 | 46 | 0 | 48 | 5 | 0 | 48 | 0 | | | % distinct signals | | 100% | 0% | 0% | 0% | 96% | 0% | 100% | 10% | 0% | 100% | 0% | | Los Pos | Pos test results | 48 | | | | | | | | | | | | | | Neg test results | 0 | | | | | | | | | | | | | | Distinct signals | | 48 | 0 | 0 | 0 | 0 | 0 | 48 | 0 | 0 | 0 | 0 | | | % distinct signals | | 100% | 0% | 0% | 0% | 0% | 0% | 100% | 0% | 0% | 0% | 0% | | High Neg | Pos test results | 6 | | | | | | | | | | | | | | Neg test results | 42 | | | | | | | | | | | | | | Distinct signals | | 6 | 0 | 48 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | | | % distinct signals | | 13% | 0% | 100% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | | Mod Neg | Pos test results | 0 | | | | | | | | | | | | | | Neg test results | 48 | | | | | | | | | | | | | | Distinct signals | | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | | | % distinct signals | | 0% | 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 | 0 | | | % distinct signals | | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | K220016 - Page 9 of 14 {9} # Reproducibility Study Eight specimens were tested in three replicates by two operators over five days across three independent sites for a total of 90 tests for each specimen. Samples were selected based on FDA cleared B. burgdorferi antibody ELISA results, including one low negative, one moderate negative, one high negative sample and one low positive, two moderate positive, one high positive sample. Final positive or negative agreement was $100\%$ for all specimens with the exception a low positive sample which produced a positive result for 77/90 replicates (positive agreement $85.5\%$ ). Similarly, a high negative sample exhibited a positive result for 2/88 replicates (negative agreement $97.8\%$ ). Table 3. Reproducibility Study Results Summary | | | | Antigens | | | | | | | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | Sample | Reactivity | Test results | VlsE | p93 | p58 | p45 | p39 | p30 | p23 | p21 | p19 | p18 | p17 | | High Pos | Poss test results | 90 | | | | | | | | | | | | | | Neg test results | 0 | | | | | | | | | | | | | | Distinct signals | | 90 | 0 | 74 | 0 | 90 | 54 | 90 | 90 | 6 | 90 | 0 | | | % distinct signals | | 100% | 0% | 82% | 0% | 100% | 60% | 100% | 100% | 7% | 100% | 0% | | Mod. Pos | Pos test results | 90 | | | | | | | | | | | | | | Neg test results | 0 | | | | | | | | | | | | | | Distinct signals | | 90 | 0 | 0 | 90 | 1 | 90 | 5 | 85 | 90 | 90 | 90 | | | % distinct signals | | 100% | 0% | 0% | 100% | 1% | 100% | 6% | 94% | 100% | 100% | 100% | | Mod Pos | Pos test results | 90 | | | | | | | | | | | | | | Neg test results | 0 | | | | | | | | | | | | | | Distinct signals | | 90 | 90 | 90 | 1 | 90 | 0 | 0 | 86 | 1 | 90 | 0 | | | % distinct signals | | 100% | 100% | 100% | 1% | 100% | 0% | 0% | 96% | 1% | 100% | 0% | | Mod Pos | Pos test results | 90 | | | | | | | | | | | | | | Neg test results | 0 | | | | | | | | | | | | | | Distinct signals | | 90 | 0 | 0 | 0 | 82 | 0 | 90 | 6 | 0 | 76 | 0 | | | % distinct signals | | 100% | 0% | 0% | 0% | 91% | 0% | 100% | 7% | 0% | 84% | 0% | | Low Pos | Pos test results | 77 | | | | | | | | | | | | | | Neg test results | 13 | | | | | | | | | | | | | | Distinct signals | | 90 | 0 | 0 | 2 | 0 | 0 | 77 | 0 | 0 | 0 | 0 | | | % distinct signals | | 100% | 0% | 0% | 2% | 0% | 0% | 86% | 0% | 0% | 0% | 0% | | High Neg | Pos test results | 2 | | | | | | | | | | | | | | Neg test results | 88 | | | | | | | | | | | | | | Distinct signals | | 2 | 0 | 89 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | | | % distinct signals | | 2% | 0% | 99% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | | Mod Neg | Pos test results | 0 | | | | | | | | | | | | | | Neg test results | 90 | | | | | | | | | | | | | | Distinct signals | | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | | | % distinct signals | | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | | Low Neg | Pos test results | 0 | | | | | | | | | | | | | | Neg test results | 90 | | | | | | | | | | | | | | Distinct signals | | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | | | % distinct signals | | 1% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | 0% | # 2. Linearity: Not applicable. K220016 - Page 10 of 14 {10} # 3. Analytical Specificity/Interference: Analytical Specificity Study: For determination of analytical specificity, 100 sera form normal blood donor individuals representing endemic and non-endemic geographic regions of the United States were tested for $B$ burgdorferi antibodies by the Borrelia All-In-One ViraChip. Table 4. Analytical Specificity Study Results Summary | | Total | Negative | Positive | % Positive | % Negative | | --- | --- | --- | --- | --- | --- | | Endemic | 50 | 46 | 4 | 8% | 92% | | Non-endemic | 50 | 50 | 0 | 0% | 100% | | All Specimens | 100 | 96 | 4 | 4% | 96% | Cross-Reactivity Study: A total of 203 sera determined to contain antibodies to other infectious disease agents were evaluated on the Borrelia All-In-One ViraChip. The results are shown in the table below. Table 5. Cross-Reactivity Study Results Summary | Disease State Sera | Total | Borrelia All-In-One ViraChip Positive | % Cross-reactivity | | --- | --- | --- | --- | | Autoimmune1 | 10 | 0 | 0% | | Babesia microti | 9 | 42 | 44.4% | | Borrelia hermsii | 6 | 0 | 0% | | Celiac disease1 | 10 | 0 | 0% | | Chlamydia trachomatis | 15 | 1 | 6.7% | | Cytomegalovirus | 11 | 33 | 27.3% | | Epstein–Barr virus | 10 | 0 | 0% | | Ehrlichia chaffeensis | 11 | 2 | 18.2% | | Fibromyalgia | 2 | 0 | 0% | | Helicobacter pylori | 10 | 0 | 0% | | Herpes simplex virus | 10 | 0 | 0% | | Influenza A | 10 | 1 | 10% | | Leptospira interrogans | 10 | 0 | 0% | | Lupus1 | 10 | 0 | 0% | | Parvovirus B19 | 10 | 0 | 0% | | Rheumatoid arthritis1 | 10 | 0 | 0% | | Rickettsia spp. | 10 | 22 | 20% | | Rubella virus | 10 | 1 | 10% | | Toxoplasma gondii | 10 | 1 | 10% | | Treponema pallidum | 10 | 0 | 0% | | Varicella zoster virus | 9 | 1 | 11.1% | Autoimmune diseases are not an infectious disease but can produce autoimmune antibodies with varied known and unknown specificity. 2Possible co-infection with $B$ burgdorferi. 2/4 positive $B$ microti samples were also positive using STTT methodology. 1/2 positie Rickettsia spp. samples were also positive by STTT methodology. $^{3}6 / 10$ samples were positive by STTT methodology K220016 - Page 11 of 14 {11} Interfering Substances: Hemolyzed, lipemic, icteric, or microbially contaminated sera should not be used for testing by the Borrelia All-In-One ViraChip. In addition the effect of elevated bilirubin and triglycerides on test results is not established. 4. Assay Reportable Range: Not applicable. 5. Traceability, Stability, Expected Values (Controls, Calibrators, or Methods): Not applicable. 6. Detection Limit: Not applicable. 7. Assay Cut-Off: The cut-off of the ViraChip test system is defined for each antigen individually by multiplying the mean intensity of the calibrator spots with an antifen specific factor. Specific antigen factors were adjusted for each antigen according to concordance with pre-characterized blood donor sera. Using Receiver Operating Characteristic analysis, the antigen specific cut-off factors were optimized for both maximum sensitivity and specificity for each antigen. ## B Comparison Studies: 1. Method Comparison with Predicate Device: Prospective Study: 113 specimens from three independent commercial clinical testing laboratories were evaluated with the Viramed Borrelia All-In-One ViraChip and comparator B. burgdorferi antibody testing following a Standard Two-Tiered Testing (STTT) methodology in which samples with positive or equivocal results from the ZEUS ELISA Borrelia VlsE1/pepC10 IgG/IgM Test System (K113397) were subsequently tested with the Borrelia B31 ViraChip IgG test kit (K163504) and Borrelia B31 ViraChip IgM test kit (K163695). The percent agreement between the Viramed Borrelia All-In-One ViraChip results and STTT results are shown below. Table 6. Prospective Study Results – Comparison with STTT | Borrelia All-In-One ViraChip | STTT Results (IgG and/or IgM) | | | | --- | --- | --- | --- | | | Positive | Negative | Total | | Positive | 41 | 16 | 57 | | Negative | 3 | 53 | 56 | | Total | 44 | 69 | 113 | Positive Percent Agreement: 93.2% (41/44) 95% CI: 81.8-97.7% Negative Percent Agreement: 76.8% (53/69) 95% CI: 65.6-85.2% K220016 - Page 12 of 14 {12} 2. Matrix Comparison: Not applicable. C Clinical Studies: 1. Clinical Sensitivity: Sensitivity Study: Staged clinically defined sera sourced from Massachusetts General Hospital were evaluated with the Viramed Biotech AG Borrelia All-In-One ViraChip and the STTT methodology described in VII.B.1 above. Results are summarized below. Table 7. Sensitivity Study Results – Comparison with STTT | | Early Lyme (N = 5) | | Disseminated Lyme (N = 19) | | Lyme Arthritis (N = 37) | | | --- | --- | --- | --- | --- | --- | --- | | | Viramed | STTT | Viramed | STTT | Viramed | STTT | | Positive | 2 | 4 | 19 | 19 | 35 | 35 | | Negative | 3 | 1 | 0 | 0 | 2 | 2 | | Sensitivity or PPA | 40% | 80% | 100% | 100% | 95% | 95% | CDC Serum Panel: A Lyme disease panel containing 90 clinically defined positive samples was obtained from the Centers for Disease Control and Prevention. These samples were evaluated with the Viramed Borrelia All-In-One ViraChip and the STTT methodology. Table 8. CDC Reference Panel Testing Results – Comparison with STTT | | Stage I (N = 60) | | Stage II (N = 10) | | Stage III (N = 20) | | Healthy Controls (N = 100) | | Disease Controls (N = 90) | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | Viramed | STTT | Viramed | STTT | Viramed | STTT | Viramed | STTT | Viramed | STTT | | Positive | 54 | 37 | 10 | 9 | 20 | 20 | 1 | 0 | 2 | 0 | | Negative | 6 | 23 | 0 | 10 | 0 | 0 | 99 | 100 | 88 | 90 | | Sensitivity or PPA | 90% | 61.7% | 100% | 90% | 100% | 100% | 99% | 100% | 98% | 100% | 2. Clinical Specificity: Not applicable. K220016 - Page 13 of 14 {13} 3. Other Clinical Supportive Data (When 1. and 2. Are Not Applicable): Not applicable. # D Clinical Cut-Off: Not applicable. # E Expected Values/Reference Range: The incidence of IgG and IgM antibodies to $B$ burgdorferi antigens in different patient populations tested by the Viramed Borrelia All-In-One ViraChip test are shown in the table below. Table 9. Expected Reactivities Among Viramed Borrelia All-In-One ViraChip Antigen Spots | | Antigens (% incidence) | | | | | | | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | Viramed Borrelia All-In-One ViraChip | VlsE | p93 | p58 | p45 | p39 | p301 | p23 | p21 | p19 | p18 | p17 | | Early Lyme Disease | 89.2% | 0.0% | 3.1% | 7.7% | 24.6% | 46.2% | 80.0% | 15.4% | 0.0% | 30.8% | 7.7% | | Cardiac Lyme | 100% | 0.0% | 0.0% | 33.3% | 66.7% | 66.7% | 66.7% | 66.7% | 0.0% | 66.7% | 0.0% | | Disseminated Lyme Disease | 100% | 0.0% | 10.5% | 31.6% | 36.8% | 63.2% | 100% | 36.8% | 0.0% | 68.4% | 31.6% | | Neurologic Lyme | 100% | 0.0% | 0.0% | 0.0% | 28.6% | 71.4% | 100% | 42.9% | 0.0% | 57.1% | 0.0% | | Prospective Study | 68.1% | 5.3% | 8.8% | 4.4% | 26.5% | 27.4% | 41.6% | 15.9% | 1.8% | 23.9% | 5.3% | | Non-Endemic Blood Donors | 1.0% | 1.0% | 0.0% | 0% | 0.0% | 1.0% | 1.0% | 0.0% | 0.0% | 1.0% | 0% | | Endemic Blood Donors | 12.0% | 0.0% | 1.0% | 0% | 1.0% | 3.0% | 4.0% | 2.0% | 0.0% | 1.0% | 2.0% | 1According to the interpretation criteria, p30 is counted only for a positive first test result (i.e, VlsE $\geq 100$ ) not an equivocal first test result. # VIII Proposed Labeling: The labeling supports the finding of substantial equivalence for this device. # IX Conclusion: The submitted information in this premarket notification is complete and supports a substantial equivalence decision. K220016 - Page 14 of 14