LYME DISEASE B. BURGDORFERI GENOGROUP 1 WESTERN BLOT IGM (WB0400M)

{0} JUN 1 1998 MRL DIAGNOSTICS K971007 510(k) Safety and Effectiveness Summary (Page 1 of 6) Applicant: MRL Diagnostics 10703 Progress Way Cypress, California 90630 Establishment Registration No: 2023365 Contact Person: Michael J. Wagner Phone: (714) 220-1900 Telefax: (714) 220-1182 E-mail: mwagner@mrlinfo.com Summary Date: May 20, 1998 Device Name: Lyme Disease B. burgdorferi Genogroup 1 Western Blot IgM Classification: Lyme Disease Borrelia burgdorferi Serological Reagents 21 CFR §866.3830 Class II Predicate Device: MRL Diagnostics Lyme Disease IFA IgM kit (K883767) 10703 PROGRESS WAY, CYPRESS CALIFORNIA 90630 800 445-0185 714 220-1900 FAX • 714 220-1182 mwagner@mrlinfo.com {1} MRL DIAGNOSTICS K971007 # 510(k) Safety and Effectiveness Summary (Page 2 of 6) ## Device Description: MRL Diagnostics separates *B. burgdorferi* proteins by polyacrylamide gel electrophoresis and electrophoretically transfers the fractionated proteins from the gel to a nitrocellulose membrane. The nitrocellulose membrane is dried and cut into strips. The antigen strips are numbered and packaged. The MRL Diagnostics Lyme Disease *B. burgdorferi* Genogroup 1 Western Blot IgM test is a two stage procedure. In the first stage, the patient sera is diluted and incubated with individual antigen strips. If antibodies to *B. burgdorferi* are present in the sera, they will bind to the Borrelia antigens immobilized on the nitrocellulose membranes. In the second stage, visualization of the bound antibodies is accomplished by incubating the blots with alkaline phosphatase—conjugated goat anti-human IgM, (Fc fragment specific) followed by the addition of substrate (BCIP/NBT) which forms a colored precipitate at each site (antigen band) where the anti-human conjugate has bound. The resulting pattern of band reactivity is then interpreted. ## Intended Use: MRL Diagnostics’ Lyme Disease *Borrelia burgdorferi* Genogroup 1 Western Blot IgM test allows the qualitative detection of IgM class antibody in human serum to individual proteins of the genogroup 1 (*B. burgdorferi* sensu stricto) of *B. burgdorferi* sensu lato. The MRL Diagnostics Lyme Disease *B. burgdorferi* Western Blot assay is intended to provide supportive evidence of infection with *B. burgdorferi*, the causative agent of Lyme disease, using serum samples which have been found positive or equivocal by another *B. burgdorferi* detection methodology (e.g. IFA or ELISA). The MRL Diagnostics Lyme Disease *B. burgdorferi* Genogroup 1 Western Blot IgM can be used alone during the acute phase (0-4 weeks of symptoms onset) of *B. burgdorferi* infection. After the acute phase, infected patients are usually found to be Western Blot positive for IgG. For persons beyond the acute phase, a positive IgM test alone is not recommended for determining active disease in patients. Therefore, both IgG and IgM Western Blots should be used after the acute phase. 10703 PROGRESS WAY, CYPRESS CALIFORNIA 90630 800 445-0185 714 220-1900 FAX • 714 220-1182 mwagner@mrlinfo.com {2} MRL DIAGNOSTICS K971007 # 510(k) Safety and Effectiveness Summary (Page 3 of 6) ## Expected Values: Three investigational sites (2 independent off-site investigators and 1 on-site investigator) studied the following 3 distinct populations: 1) a **Lyme Disease population** (n=181) consisting of serum samples from patients with EM, a late stage symptom per the CDC case definition and *B. burgdorferi* seropositive, or *B. burgdorferi* culture positive; 2) a **Normal population** (n=325) consisting of serum samples from donors with no known history or symptoms of Lyme Disease (including persons from Lyme disease endemic and non-endemic areas), and; 3) a **Cross-reactivity population** (n=281) consisting of serum samples from patients with potentially cross-reactive conditions or infections. The frequency of criteria bands observed in the three populations are described in the following table: | Disease State | n | %Pos | Any Band | Criteria Bands (kDa) | | | | --- | --- | --- | --- | --- | --- | --- | | | | | | 23 | 39 | 41 | | Lyme Disease (By Disease Stage) (n=181) | | | | | | | | Lyme Stage I | 72 | 69% | 86% | 71% | 49% | 55% | | Lyme Stage II | 42 | 66% | 83% | 59% | 56% | 76% | | Lyme Stage III | 67 | 30% | 70% | 27% | 31% | 42% | | Lyme Disease (By Months Post Infection Onset) (n=181) | | | | | | | | < 1 month | 40 | 78% | 80% | 73% | 53% | 80% | | 1 to 2 months | 25 | 72% | 92% | 72% | 60% | 76% | | 3 to 12 months | 63 | 48% | 71% | 45% | 39% | 58% | | > 12 months | 53 | 34% | 81% | 34% | 36% | 49% | | Normal Population (n=325) | | | | | | | | Endemic | 81 | 0% | 7% | 1% | 0% | 2% | | Non-Endemic | 244 | 2% | 23% | 11% | 2% | 9% | | Potential Cross-reactives (n=281) | | | | | | | | Spirochetal | 104 | 3% | 40% | 19% | 5% | 12% | | Auto-Immune | 22 | 18% | 55% | 41% | 0% | 23% | | Tick Borne | 21 | 24% | 38% | 29% | 5% | 29% | | Muscular Skeletal | 33 | 3% | 24% | 9% | 0% | 6% | | Neurologic | 27 | 4% | 41% | 7% | 0% | 22% | | Viral | 32 | 0% | 13% | 3% | 0% | 9% | | Symptomology | 15 | 0% | 27% | 13% | 0% | 13% | | Misc. Disease | 27 | 4% | 15% | 4% | 4% | 11% | 10703 PROGRESS WAY, CYPRESS CALIFORNIA 90630 800 445-0185 714 220-1900 FAX • 714 220-1182 mwagner@mrlinfo.com {3} MRL DIAGNOSTICS K971007 510(k) Safety and Effectiveness Summary (Page 4 of 6) ## Sensitivity: To determine assay Sensitivity, two independent off-site investigators (Investigational Sites 1 and 2) and one on-site investigator (Investigational Site 3), assayed well characterized Lyme disease positive patient sera (n=176) from patients (n=155) with EM, a late stage symptom per the CDC case definition and B. burgdorferi seropositive, or B. burgdorferi culture positive. Investigational Sites 1 and 2 used retrospective serum samples from their own well-characterized serum banks, and Investigational Site 3 used a retrospective well characterized serum panel supplied by the CDC. The CDC panel results are presented as a means to convey further information on the performance of this assay with a masked, characterized serum panel. This does not imply an endorsement of the assay by the CDC. For the CDC panel, sensitivity results are provided in the following table for 1) the IgM Western Blot, and 2) the number of samples which were IgM negative but IgG positive by MRL Diagnostics' B. burgdorferi Western blots ("IgM Negative &amp; IgG Positive"): | Months Post Infection Onset | IgG Sensitivity | IgG 95% CI | IgG Negative & IgM Positive | | --- | --- | --- | --- | | <1 | 100% (4/4) | 40 to 100% | 0 | | 1 to 2 | 79% (7/9) | 40 to 97% | 1 | | 3 to 6 | 75% (12/16) | 48 to 97% | 2 | | >6 | 36% (4/11) | 11 to 69% | 6 | | Overall Sensitivity | 66% (27/40) | 51 to 84% | 9 | For all three sites, sensitivity results are provided in the following table for 1) the IgM Western Blot and 2) the number of samples which were IgM negative but IgG positive by MRL Diagnostics' B. burgdorferi Western blots ("IgM Negative &amp; IgG Positive"): | Months Post Infection Onset | IgM Sensitivity | IgM 95% CI | IgG Positive & IgM Negative | | --- | --- | --- | --- | | <1 | 77% (30/39) | 61 to 89% | 1 | | 1 to 2 | 72% (18/25) | 51 to 88% | 4 | | 3 to 6 | 63% (25/40) | 46 to 77% | 7 | | >6 | 32% (23/71) | 22 to 45% | 29 | | Unspecified | 0% (0/1) | | 1 | | Overall Sensitivity | 55% (97/176) | 47 to 63% | 42 | Eighteen patients were drawn more than once, i.e., three patients were drawn three times and the other patients twice. One of those patients was drawn at 99 and 105 months post onset, and both draws were found IgG positive IgM negative. The other seventeen patients were initially drawn less than one month post onset, and the results for those multiple draws are summarized in the following table: | | Initial Draw | Subsequent Draw Sensitivity | | | | --- | --- | --- | --- | --- | | | Sensitivity | <1 Mo. | 1-2 Mo. | 3-6 Mo. | | | 76% (13/17) | 100% (1/1) | 90% (9/10) | 67% (6/9) | 10703 PROGRESS WAY, CYPRESS CALIFORNIA 90630 800 445-0185 714 220-1900 FAX • 714 220-1182 mwagner@mrlinfo.com {4} MRL DIAGNOSTICS K971007 # 510(k) Safety and Effectiveness Summary (Page 5 of 6) ## Specificity To determine assay Specificity, Investigational Sites 1, 2, and 3 assayed 606 sera consisting of 325 serum samples from persons with no known history of Lyme disease (from endemic and non-endemic areas) (Investigational Sites 1, 2, and 3) and 284 disease state serum samples from potentially cross-reactive diseases (including diseases likely to produce similar clinical presentation) (Investigational Sites 1 and 3). Where specimen quantities were sufficient, the investigators assayed each specimen on both the IgG and the IgM Western Blots. For persons with no known history of Lyme disease (Normals), specificity results are provided in the following table for 1) the IgM Western Blot and 2) the number of samples which were IgM negative but IgG positive by MRL Diagnostics’ *B. burgdorferi* Western blots (“IgM Negative &amp; IgG Positive”): | Condition | IgM Positivity | IgG Positive & IgM Negative | | --- | --- | --- | | **Normal Population (n=325)** | | | | Endemic | 0% (0/81) | 0 | | Non-endemic | 2%(5/244) | 0 | | Overall Normal Positivity | 2%(5/325) | 0 | | Condition | IgM Cross-reactivity | IgG Positive & IgM Negative | | **Spirochetal Disease Population (n=104)** | | | | Syphilis | 1%(1/74) | 0 | | Periodontal | 10%(2/20) | 0 | | Leptospirosis | 0%(0/10) | 0 | | **Auto Immune Disease Population (n=22)** | | | | RF | 20%(2/10) | 0 | | ANA | 18%(2/11) | 0 | | SLE (Lupus) | 0% (0/1) | 0 | | **Tickborne Disease Population (n=21)** | | | | Rickettsia | 0% (0/9) | 0 | | E. chaffeensis | 63% (5/8) | 0 | | HGE | 0% (0/3) | 0 | | Tickborne Relapsing | 0% (0/1) | 0 | | **Muscular Skeletal Disease (n=33)** | | | | Arthritis | 0%(0/15) | 1 | | Arthralgia | 9%(1/11) | 1 | | JRA | 0% (0/4) | 0 | | Misc. Muscular | 0% (0/3) | 0 | | **Misc. Neurologic Disease (n=27)** | 4%(1/27) | 1 | | **Viral Disease Population (n=36*)** | | | | EBV* | 0%(0/13) | 0 | | CMV* | 0%(0/11) | 0 | | HSV-1/2* | 0% (0/2) | 0 | | HIV | 0%(0/10) | 0 | | **Misc. Disease/Similar Symptoms (n=42)** | | | | Fatigue | 0%(0/15) | 0 | | Misc. | 4%(1/27) | 0 | * includes 2 sera with antibody response to EBV, CMV, and HSV-1/2 10703 PROGRESS WAY, CYPRESS CALIFORNIA 90630 800 445-0185 714 220-1900 FAX • 714 220-1182 mwagner@mrlinfo.com {5} MRL DIAGNOSTICS K971007 # 510(k) Safety and Effectiveness Summary (Page 6 of 6) ## Reproducibility **Intra-laboratory Reproducibility** was assessed by assaying 10 serum samples once per day on 3 days using a single MRL Diagnostics Western Blot Genogroup 1 Strip lot. All 3 runs yielded identical screening interpretations for all 10 selected sera. Overall, the 3 criteria bands yielded identical interpretations on 89 of 90 (99%) criteria band readings. **Inter-lot Reproducibility** was assessed using 10 serum samples on 3 lots of MRL Diagnostics Western Blot Genogroup 1 Strips. All 3 lots produced identical screening interpretations for all 10 selected sera. Overall, the 3 criteria bands yielded identical interpretations on 89 of 90 (99%) criteria band readings. **Inter-reader Reproducibility** was assessed by assaying 18 patient sera with a single lot of strips. The sera were assayed at the three Investigational Sites, and read by two readers at each site. Overall, interpretation was identical 94% (51/54), and criteria band readings were identical 96% (155/162). **Inter-site Reproducibility** was assessed by assaying 18 patient sera with a single lot of strips. The sera were assayed at the three Investigational Sites, and read by two readers at each site. Overall, interpretation was identical 95% (97/108), and criteria band readings were identical 96% (311/324). The following table summarizes the reproducibility studies by interpretation and band reproducibility. | Study | n | Interp | 23 | 39 | 41 | | --- | --- | --- | --- | --- | --- | | Intra-lab | 10 | 100% | 100% | 90% | 100% | | Inter-lot | 10 | 100% | 100% | 100% | 90% | | Inter-reader | 18 | 96% | 96% | 94% | 96% | | Inter-site | 18 | 95% | 98% | 94% | 96% | 10703 PROGRESS WAY, CYPRESS CALIFORNIA 90630 800 445-0185 714 220-1900 FAX • 714 220-1182 mwagner@mrlinfo.com {6} DEPARTMENT OF HEALTH &amp; HUMAN SERVICES Public Health Service Food and Drug Administration 2098 Gaither Road Rockville MD 20850 JUN 1 1998 Michael J. Wagner Regulatory Affairs Specialist MRL Diagnostics 10703 Progress Way Cypress, CA 90630 Re: K971007 Trade Name: Lyme Disease B. burgdorferi Genogroup 1 Western Blot IgM Regulatory Class: II Product Code: LSR Dated: March 23, 1998 Received: March 25, 1998 Dear Mr. Wagner: We have reviewed your Section 510(k) notification of intent to market the device referenced above and we 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). 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 either class II (Special Controls) or class III (Premarket Approval), it may be subject to such additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 895. A substantially equivalent determination assumes compliance with the Current Good Manufacturing Practice requirements, as set forth in the Quality System Regulation (QS) for Medical Devices: General regulation (21 CFR Part 820) and that, through periodic QS inspections, the Food and Drug Administration (FDA) will verify such assumptions. Failure to comply with the GMP regulation may result in regulatory action. In addition, FDA may publish further announcements concerning your device in the Federal Register. Please note: this response to your premarket notification submission does not affect any obligation you might have under sections 531 through 542 of the Act for devices under the Electronic Product Radiation Control provisions, or other Federal laws or regulations. {7} Page 2 Under the Clinical Laboratory Improvement Amendments of 1988 (CLIA-88), this device may require a CLIA complexity categorization. To determine if it does, you should contact the Centers for Disease Control and Prevention (CDC) at (770)488-7655. This letter will allow you to begin marketing your device as described in your 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 Part 801 and additionally 809.10 for *in vitro* diagnostic devices), please contact the Office of Compliance at (301) 594-4588. Additionally, for questions on the promotion and advertising of your device, please contact the Office of Compliance at (301) 594-4639. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers Assistance at its toll free number (800) 638-2041 or at (301) 443-6597 or at its internet address "http://www.fda.gov/cdrh/dsmamain.html" Sincerely yours, Steven I. Gutman, M.D., M.B.A. Director Division of Clinical Laboratory Devices Office of Device Evaluation Center for Devices and Radiological Health Enclosure {8} Page 1 of 1 510(k) Number (if known): K971007 Device Name: Lyme Disease B. burgdorferi Genogroup 1 Western Blot IgM Indications for Use: MRL Diagnostics’ Lyme Disease Borrelia burgdorferi Genogroup 1 Western Blot IgM test allows the qualitative detection of IgM class antibody in human serum to individual proteins of the genogroup 1 (B. burgdorferi sensu stricto) of B. burgdorferi sensu lato. The MRL Diagnostics Lyme Disease B. burgdorferi Western Blot assay is intended to provide supportive evidence of infection with B. burgdorferi, the causative agent of Lyme disease, using serum samples which have been found positive or equivocal by another B. burgdorferi detection methodology (e.g. IFA or ELISA). The MRL Diagnostics Lyme Disease B. burgdorferi Genogroup 1 Western Blot IgM can be used alone during the acute phase (0-4 weeks of symptoms onset) of B. burgdorferi infection. After the acute phase, infected patients are usually found to be Western Blot positive for IgG. For persons beyond the acute phase, a positive IgM test alone is not recommended for determining active disease in patients. Therefore, both IgG and IgM Western Blots should be used after the acute phase. PLEASE DO NOT WRITE BELOW THIS LINE - CONTINUE ON ANOTHER PAGE IF NEEDED) Concurrence of CDRH, Office of Device Evaluation (ODE) Woody Dubais (Division Sign Off) Division of Clinical Laboratory Devices 510(k) Number K971007 Prescription Use ☑ (Per 21 CFR 801.109) OR Over-The-Counter Use ☐ (Optional Format 1-2-96)