(90 days)
The BioPlex 2200 Lyme Total kit is a multiplex flow immunoassay intended for the qualitative detection of total (lgM/IgG) antibodies to Borrelia burgdorferi in human serum and plasma (EDTA, heparin). This assay should be used to test patients with a history and/ or symptoms of infection with B. burgdorferi. The BioPlex 2200 Lyme Total assay is intended for use with the Bio-Rad BioPlex 2200 System. All reactive and equivocal specimens should be tested with a second tier test such as Lyme IgM and IgG Western blot assays. Positive second tier results are supportive evidence of infection with B. burgdorferi. Diagnosis of Lyme borreliosis should be made based on the presence of B. burgdorferi antibodies, history, symptoms, and other laboratory data. Non-reactive first tier or negative second tier results should not be used to exclude borreliosis.
BioPlex 2200 Lyme Total kit includes the following components:
- One (1) 10 mL vial of Bead Set , containing dyed beads coated with recombinant p58, OspC ● type B (OspCB) and synthetic peptide FVlsE (consisting of FlaB and VlsE sequences), an Internal Standard bead (ISB) and a Serum Verification bead (SVB) in MOPS (3-[N-Morpholino] propane sulfonic acid) buffer containing bovine proteins with protein stabilizers. BND (5-bromo-5-nitro-1,3-dioxan) (≤ 0.1%), ProClin 300 (≤ 0.3%), sodium benzoate (≤ 0.1%) and sodium azide (< 0.1%) are added as preservatives.
- One (1) 5 mL vial of Conjugate, containing phycoerythrin conjugated murine monoclonal anti-. human IgG, and murine monoclonal anti-human IgM, and phycoerythrin conjugated murine monoclonal anti-human FXIII antibody in phosphate buffer, supplemented with murine and bovine protein stabilizers. ProClin 300 (≤ 0.3%) and sodium azide (< 0.1%) are added as preservatives.
- One (1) 10 mL vial of Sample Diluent, containing bovine and murine proteins in ● triethanolamine buffer. ProClin 300 (≤ 0.3%), sodium benzoate (≤ 0.1%) and sodium azide (< 0.1%) are added as preservatives.
Additional materials required but not supplied include BioPlex 2200 Sheath Fluid containing Phosphate Buffered Saline (PBS) with ProClin 300 (0.03%) and sodium azide (<0.1%) added as preservatives; and the BioPlex 2200 Wash Solution containing Phosphate Buffered Saline (PBS) and Tween 20 with ProClin 300 (0.03%) and sodium azide (<0.1%) added as preservatives.
The document provided is a 510(k) Summary for the BioPlex 2200 Lyme Total kit, which is an in vitro diagnostic (IVD) device for the qualitative detection of total (IgM/IgG) antibodies to Borrelia burgdorferi. It is not an AI/ML-driven device, nor does it involve human readers interpreting images. Therefore, many of the requested criteria in your prompt (such as human readers, AI assistance, expert consensus for ground truth, adjudication methods, or sample size for training sets) are not applicable to this type of device.
However, I can extract information related to the acceptance criteria and performance study from the provided document as it pertains to an IVD device.
Key takeaway: This document describes a traditional in-vitro diagnostic device, not an AI/ML system for diagnostic imaging. Therefore, concepts like "human readers improve with AI vs without AI assistance" or "standalone (i.e. algorithm only without human-in-the-loop performance)" are irrelevant. The "acceptance criteria" for this IVD device would typically refer to predefined performance specifications agreed upon for regulatory clearance.
Here's the relevant information based on the document provided:
1. Table of Acceptance Criteria and Reported Device Performance
For IVD devices, acceptance criteria are generally established for various performance characteristics like sensitivity, specificity, precision, cross-reactivity, and interference. The document presents the results of these studies, and the implicit acceptance is that these results demonstrate substantial equivalence to the predicate device and are clinically acceptable. The performance is compared against a commercial Lyme IgM/IgG immunoassay or a CDC reference panel.
| Performance Metric | Acceptance Criteria (Implicit from Study Design/Context) | Reported Device Performance (BioPlex 2200 Lyme Total) | Predicate Device / Commercial Assay Performance (for comparison) |
|---|---|---|---|
| Clinical Sensitivity (Prospective Samples) | (Expected to be comparable to or better than predicate) | PPA = 70.1% (68/97) (60.0 - 79.0% CI) when compared to predicate's positive/equivocal results. | Commercial Lyme IgM/IgG Immunoassay: Pos: 88, Eqv: 9, Neg: 695 |
| Clinical Specificity (Prospective Samples) | (Expected to be comparable to or better than predicate) | NPA = 95.1% (661/695) (93.2 - 96.6% CI) when compared to predicate's negative results. | Commercial Lyme IgM/IgG Immunoassay: Pos: 88, Eqv: 9, Neg: 695 |
| Sensitivity by Disease Stage (Archived Samples) | (Expected performance across stages) | Acute: 69.4% (50/72)Convalescent: 61.5% (16/26)Late: 85.7% (6/7)Total: 68.6% (72/105) | Commercial Lyme IgM/IgG Immunoassay:Acute: 58.3% (42/72)Convalescent: 69.2% (18/26)Late: 71.4% (5/7)Total: 61.9% (65/105) |
| Precision (Within Run / Total %CV) | (Low variability expected) | Within Run: 2.7% - 6.1%Total: 4.0% - 6.8% (for various AI levels) | N/A (Internal study on device's own performance) |
| Reproducibility (Total %CV) | (Low variability across sites/days) | Total: 5.1% - 8.1% (for various AI levels) | N/A (Internal study on device's own performance) |
| Analytical Specificity (Asymptomatic Samples) | (High percentage of negatives expected) | Asymptomatic Non-Endemic: 96.4% NegativesAsymptomatic Endemic: 96.4% Negatives | Commercial Lyme IgM/IgG Immunoassay:Non-Endemic: 95.4% NegativesEndemic: 94.8% Negatives |
| Cross-Reactivity (% Negative Agreement) | (High % Agreement, low false positives with other conditions) | 91% - 100% for various cross-reactants (e.g., EBV: 92%, Syphilis: 98%) | N/A (compared to Commercial Lyme Immunoassay's negative results) |
| Matrix Comparison (Correlation r) | (High correlation between serum and plasma) | K2-EDTA vs. Serum: r=0.960K3-EDTA vs. Serum: r=0.951Sodium Heparin vs. Serum: r=0.970Lithium Heparin vs. Serum: r=0.981 | N/A |
| CDC Panel Performance | (High agreement with CDC characterized samples) | Clinical Diagnosis Agreement: Acute 84.6%, Convalescent 93.5%, Late 100%, Look-alike 96.7%, Healthy 97.0% | Comparison against another commercial assay also performed and reported. |
2. Sample Sizes Used for the Test Set and Data Provenance
- Clinical Method Comparison Study (Test Set): 792 prospectively collected serum samples from patients submitted for Lyme disease testing.
- Data Provenance: "various geographically distinct locations in the U.S." (retrospective or prospective is not explicitly stated for this part, but implies prospective collection for "patients submitted for Lyme disease testing"). The samples were evaluated at "three (3) U.S. clinical testing sites."
- Sensitivity Study (Test Set): 105 clinically characterized samples from an archived collection.
- Data Provenance: Not specified for geographical origin beyond "archived collection."
- Analytical Specificity Study (Test Set): 836 samples from asymptomatic individuals from endemic regions (416 asymptomatic non-endemic, 420 asymptomatic endemic). These were "obtained from an asymptomatic population and excluded pre-screened blood donors."
- Data Provenance: Not explicitly stated (e.g., country of origin), implies collected from relevant populations.
- Cross-Reactivity Study (Test Set): Varies per cross-reactant, e.g., 13 samples for ANA, 49 for EBV, 48 for Syphilis, etc. (Total N for this section is sum of all individual cross-reactant groups). Samples were "known to be positive for each cross-reactant" and "pre-tested by a commercially available Lyme assay and only those that tested negative by the commercially available assay were further tested."
- Precision Study: 80 replicates per panel member (6 panel members evaluated).
- Reproducibility Study: 120 total replicates per panel member (5 panel members evaluated), tested at 3 clinical trial sites.
- Matrix Comparison: Minimum of 40 sets of paired serum and plasma samples, total N for comparison ranged from 74 to 78 pairs.
- CDC Panel: 280 positive and negative specimens from the Centers for Disease Control and Prevention (CDC).
- Data Provenance: CDC (implies U.S. origin).
3. Number of Experts Used to Establish Ground Truth for the Test Set and Their Qualifications
Not Applicable / Not Provided for this IVD device. The device measures antibodies. The "ground truth" for clinical studies is based on patient clinical history, symptoms, and other laboratory data, or classification by a reference panel (like the CDC panel) or comparison to an already cleared predicate device. There is no mention of "experts" in the context of image interpretation or similar tasks requiring human consensus.
4. Adjudication Method for the Test Set
Not Applicable / Not Provided for this IVD device. Adjudication methods like "2+1, 3+1" are typically used for interpreting ambiguous cases or discordances in human reader studies, often in the context of imaging. For this IVD device, results are quantitative (Antibody Index) and then categorized into discrete "Reactive," "Equivocal," or "Non-Reactive" bins based on defined cut-offs. Discords between the new device and the predicate or clinical diagnosis are reported, but there's no mention of an adjudication process by human experts to resolve these, rather they are statistical comparisons. For equivocal results, the assay states they "should be tested with a second tier test such as Lyme IgM and IgG Western blot assays."
5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance
Not Applicable. This document describes an IVD antibody detection kit, not an AI-assisted imaging device or a device involving human readers.
6. If a Standalone (i.e. algorithm only without human-in-the-loop performance) was done
The BioPlex 2200 Lyme Total kit is a standalone device in the sense that it performs the assay and provides quantitative results (Antibody Index) which are then categorized. There isn't a "human-in-the-loop" performing the primary measurement or interpretation in the way one would for an AI-assisted diagnostic imaging system. The device itself produces the raw data and converts it into the final qualitative result (Reactive, Equivocal, Non-Reactive) based on internal algorithms and calibrated cutoffs. Thus, all the performance data presented (sensitivity, specificity, precision, etc.) represents its standalone performance as an automated IVD assay.
7. The Type of Ground Truth Used
The "ground truth" for this IVD device's performance studies varies:
- Clinical Diagnosis: For the "Sensitivity Study" and "CDC Panel," samples were "clinically characterized" by disease stage (e.g., Acute, Convalescent, Late Lyme) or defined categories (e.g., look-alike diseases, healthy controls).
- Predicate Device Comparison: In the "Method Comparison Study," the new device's qualitative results were compared against an existing "Commercial Lyme IgM/IgG Immunoassay" (the predicate device). This serves as a reference for substantial equivalence.
- Second-Tier Testing (Western Blot): For reactive and equivocal results from both the new device and predicate, a "FDA-cleared IgG and IgM Western blot assays" was used as a confirmatory "second tier test." This provides a more definitive ground truth for those specific samples implicated as positive or equivocal by initial screening assays.
- Archived Collections/Known Status: For the precision, reproducibility, analytical specificity, interfering substances, and cross-reactivity studies, samples with pre-defined characteristics (e.g., specific concentrations, known disease states, known absence of target analyte) were used.
8. The Sample Size for the Training Set
Not Applicable / Not Provided. This document does not describe the development of an AI/ML algorithm that requires a specific training set. The device is a traditional immunoassay kit. The "assay cut-off" (similar to a classification threshold in ML) was determined using "1,372 normal sera" and was "retrospectively selected around the 98th percentile." This cohort was used to define the operational cut-off rather than "training" an algorithm in the AI/ML sense.
9. How the Ground Truth for the Training Set Was Established
Not Applicable. As above, there is no AI/ML training set. The assay cut-off was established using 1,372 normal sera, identifying a relative fluorescence intensity (RFI) corresponding to the 98th percentile to set the 1.0 AI cut-off. This was then "subsequently evaluated for performance against clinically diagnosed samples."
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Image /page/0/Picture/0 description: The image contains 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 consists of the letters "FDA" in a blue square, followed by the words "U.S. FOOD & DRUG" in blue, and the word "ADMINISTRATION" in a smaller font below.
March 12, 2019
Bio-Rad Laboratories Arlene Carillo RA Manager 5500 East Second Street Benicia, California 94510
Re: K183446
Trade/Device Name: BioPlex 2200 Lyme Total Regulation Number: 21 CFR 866.3830 Regulation Name: Treponema pallidum treponemal test reagents Regulatory Class: Class II Product Code: LSR Dated: December 11, 2018 Received: December 12, 2018
Dear Arlene Carillo:
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 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
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- for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.html; good manufacturing practice requirements as set forth in the quality systems (QS) 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 http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm.
For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). 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 (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely,
for
Uwe Scherf, Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
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BioPlex 2200 Lyme Total 510(k) Summary
Bio-Rad Laboratories hereby submits this 510(k) in accordance with the requirements of SMDA 1990 and 21 CFR 807.92. This summary of 510(k) safety and effectiveness information provides detail as a basis for a determination of substantial equivalence for the BioPlex 2200 Lyme Total kit.
510(k) Number: K183446
Summary Preparation Date: December 11, 2018
Applicant: Bio-Rad Laboratories
Contact: Arlene Carillo Regulatory Affairs Manager 5500 East Second Street Benicia, CA 94510
Measurand: Anti-Borrelia burgdorferi (IgM and IgG) antibodies
Type of Test: Quantitative multiplexed flow immunoassay
Proprietary and Established Names:
BioPlex 2200 Lyme Total
Regulatory Information:
-
Regulation section: 21 CFR 866.3830; Treponema pallidum treponemal test reagents
-
Classification: Class II
-
- Product code: LSR; Reagent, Borrelia Serological Reagent
-
- Panel: Microbiology (83)
Intended Use:
Intended use(s):
The BioPlex 2200 Lyme Total kit is a multiplex flow immunoassay intended for the qualitative detection of total (lgM/IgG) antibodies to Borrelia burgdorferi in human serum and plasma (EDTA, heparin). This assay should be used to test patients with a history and/ or symptoms of infection with B. burgdorferi. The BioPlex 2200 Lyme Total assay is intended for use with the Bio-Rad BioPlex 2200 System. All reactive and equivocal specimens should be tested with a second tier test such as Lyme IgM and IgG Western blot assays. Positive second tier results are
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supportive evidence of infection with B. burgdorferi. Diagnosis of Lyme borreliosis should be made based on the presence of B. burgdorferi antibodies, history, symptoms, and other laboratory data. Non-reactive first tier or negative second tier results should not be used to exclude borreliosis.
Device Description:
BioPlex 2200 Lyme Total kit includes the following components:
- One (1) 10 mL vial of Bead Set , containing dyed beads coated with recombinant p58, OspC ● type B (OspCB) and synthetic peptide FVlsE (consisting of FlaB and VlsE sequences), an Internal Standard bead (ISB) and a Serum Verification bead (SVB) in MOPS (3-[N-Morpholino] propane sulfonic acid) buffer containing bovine proteins with protein stabilizers. BND (5-bromo-5-nitro-1,3-dioxan) (≤ 0.1%), ProClin 300 (≤ 0.3%), sodium benzoate (≤ 0.1%) and sodium azide (< 0.1%) are added as preservatives.
- One (1) 5 mL vial of Conjugate, containing phycoerythrin conjugated murine monoclonal anti-. human IgG, and murine monoclonal anti-human IgM, and phycoerythrin conjugated murine monoclonal anti-human FXIII antibody in phosphate buffer, supplemented with murine and bovine protein stabilizers. ProClin 300 (≤ 0.3%) and sodium azide (< 0.1%) are added as preservatives.
- One (1) 10 mL vial of Sample Diluent, containing bovine and murine proteins in ● triethanolamine buffer. ProClin 300 (≤ 0.3%), sodium benzoate (≤ 0.1%) and sodium azide (< 0.1%) are added as preservatives.
Additional materials required but not supplied include BioPlex 2200 Sheath Fluid containing Phosphate Buffered Saline (PBS) with ProClin 300 (0.03%) and sodium azide (<0.1%) added as preservatives; and the BioPlex 2200 Wash Solution containing Phosphate Buffered Saline (PBS) and Tween 20 with ProClin 300 (0.03%) and sodium azide (<0.1%) added as preservatives.
Substantial Equivalence Information:
-
- Predicate device name(s): Immunetics C6 B. burgdorferi (Lyme) ELISA Kit, K003754
-
- Comparison with predicate:
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| Device Similarities | ||
|---|---|---|
| Characteristics | New DeviceBioPlex 2200 Lyme Total | Predicate DeviceC6 B. burgdorferi (Lyme) ELISA Kit |
| Intended Use | The BioPlex 2200 Lyme Total kit is a multiplex flow immunoassay intended for the qualitative detection of total (IgM/IgG) antibodies to Borrelia burgdorferi in human serum and plasma (EDTA, heparin). This assay should be used to test patients with history and/or symptoms of infection with B. burgdorferi . The BioPlex 2200 Lyme Total assay is intended for use with the Bio-Rad BioPlex 2200 System. All reactive and equivocal specimens should be tested with a second tier test such as Lyme IgM and IgG Western blot assays. Positive second tier results are supportive evidence of infection with B. burgdorferi . Diagnosis of Lyme borreliosis should be made based on the presence of B. burgdorferi antibodies, history, symptoms, and other laboratory data. Non-reactive first tier or negative second tier results should not be used to exclude borreliosis. | The Immunetics C6 B. burgdorferi (Lyme) ELISA Kit is intended for use in the presumptive detection of IgG and IgM antibodies to B. burgdorferi in human serum. The assay should be used only on samples from patients with clinical history, signs or symptoms consistent with B. burgdorferi infection, including individuals who have received the licensed recombinant OspA Lyme disease vaccine (Lymerix). Positive or equivocal results should be supplemented by testing with a standardized Western Blot (second step) method. Positive Western Blot results provide evidence for exposure to or infection with B. burgdorferi . The diagnosis of Lyme disease must be made based on history, signs (such as erythema migrans), symptoms, and other laboratory data, in addition to the presence of antibodies to B. burgdorferi . Negative results (either first or second step) should not be used to exclude Lyme disease. |
| MeasuredAnalyte | Anti- Borrelia burgdorferi (IgM and IgG) antibodies | Same |
| Assay Type | Qualitative | Qualitative |
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| Characteristics | Device Differences | |
|---|---|---|
| New DeviceBioPlex 2200 Lyme Total | Predicate DeviceImmunetics C6 B. burgdorferi(Lyme) ELISA Kit, K003754 | |
| Assay Technology | Automated multiplex flow competitiveimmunoassay | ELISA (enzyme-linkedimmunosorbent assay) |
| Antigen | Recombinant p58, OspC type B (OspCB)and synthetic peptide FVIsE | Synthetic peptide (C6 peptide) |
| Signal Detection | Fluorescence | Optical density readings fromSpectrophotometer |
| Solid Phase | Antigen-coated paramagnetic microbeads | Antigen-coated microwell |
| Conjugate | Murine monoclonal anti-human IgG, murinemonoclonal anti-human IgM andphycoerythrin conjugated murine monoclonalanti-human FXIII antibody | Horseradish peroxidase-conjugated (HRP) goat anti-human IgG/IgM conjugate |
| Sample Size | 5 μL | 10 μL |
| SampleHandling/Process | Automated | Manual |
| Unit of Measure | Antibody Index (AI) | Lyme Index (LI) |
| Sample Matrix | Serum and plasma | Serum |
| Instrumentation | Bio-Rad BioPlex 2200 System | None (Manual) |
Test Principle: The BioPlex 2200 Lyme Total kit employs fluoromagnetic, dyed beads which are coated with recombinant p58, OspCB or synthetic peptide FVIsE. Unique fluorescent signatures identify the presence of total (IgM/IgG) antibodies to B. burgdorferi in a two-step assay format.
The BioPlex 2200 System combines an aliquot of patient sample diluent, and bead reagent in a reaction vessel. The mixture is incubated at 37°C. After a wash cycle, a mixture of murine monoclonal anti-human IgG and murine monoclonal anti-human IgM antibody conjugated to phycoerythrin (PE) is added to the dyed beads, and this mixture is incubated at 37°C. The excess conjugate is removed in another wash cycle and the beads are re-suspended in sheath fluid. The bead mixture then passes through the detector. The identity of the dyed beads is determined by the fluorescence of the dyes, and the amount of antibody captured by the antigen is determined by the fluorescence of the attached PE. Raw data is calculated in relative fluorescence intensity (RFI).
Two additional dyed beads, an Internal Standard Bead (ISB) and a Serum Verification Bead (SVB), are present in each reaction mixture to verify detector response and the addition of serum or plasma to the reaction vessel respectively. Refer to the BioPlex 2200 System Operation Manual for more information.
The system is calibrated using a set of three (3) distinct calibrator vials supplied separately by Bio-Rad Laboratories. One vial contains negative sample, and the other two vials contain human B. burgdorferi sensu stricto antibodies, which are used for qualitative calibration of the assays. The qualitative results
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are expressed in an antibody index (AI). The assay results are reported as nonreactive, equivocal or reactive.
Performance Characteristics
Non-Clinical Studies
Assay Cut-Off: A total of 1.372 normal sera were used to determine the percentile cut-off in increments of 1/1000th for each analyte. Based on a retrospective selection of around the 98th percentile, the relative fluorescence intensity (RFI) corresponding to the chosen cutoff was assigned a value of 1.0 AI, and subsequently evaluated for performance against clinically diagnosed samples. The statistics show that the 98th percentile cutoff meets the minimum agreement specifications while maximizing the clinical specificity. The assay employs an equivocal zone that brackets the cut-off (0.9-1.0 AI), with samples ≥1.1 AI reported as reactive and ≤0.8 AI reported as non-reactive.
Precision Study: Precision testing of the BioPlex 2200 Lyme Total kit was performed in accordance with the Clinical and Laboratory Standards Institute (CLSI) EP5-A3 guideline. The serum samples were tested in duplicate, two runs per day, over 20 days (2 replicates x 2 runs x 20 days = 80 replicates per panel member) using one lot of the BioPlex 2200 Lyme Total Reagent Pack, Calibrator Set and Control Set. The results are summarized in the table below.
| Within Run(Repeatability) | BetweenRun | BetweenDay | Total | ||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SampleType | SampleDescription | N | Mean(AI) | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Serum | HighNegative 1 | 80 | 0.7 | 0.032 | 4.5% | 0.000 | 0.0% | 0.015 | 2.1% | 0.035 | 4.9% |
| HighNegative 2 | 80 | 0.9 | 0.050 | 5.4% | 0.000 | 0.0% | 0.010 | 1.1% | 0.051 | 5.5% | |
| Cut-Off | 80 | 1.0 | 0.040 | 3.9% | 0.000 | 0.0% | 0.030 | 2.8% | 0.050 | 4.8% | |
| LowPositive | 80 | 1.5 | 0.040 | 2.7% | 0.032 | 2.1% | 0.031 | 2.1% | 0.060 | 4.0% | |
| Mid Positive | 80 | 2.8 | 0.081 | 2.9% | 0.071 | 2.5% | 0.039 | 1.4% | 0.114 | 4.1% | |
| HighPositive | 80 | 4.7 | 0.288 | 6.1% | 0.000 | 0.0% | 0.139 | 3.0% | 0.320 | 6.8% |
BioPlex 2200 Lyme Total Precision
Reproducibility Study: To assess reproducibility of the BioPlex 2200 Lyme Total kit, a reproducibility panel was prepared at Bio-Rad Laboratories. The panel contained members with varying levels of antibodies to the analytes in the BioPlex 2200 Lyme Total kit, and a positive control (antibody reactive for all analytes). Reproducibility testing was performed at 3 clinical trial sites. One lot of BioPlex 2200 Lyme Total Reagent Packs, BioPlex 2200 Lyme Total Calibrator Sets and BioPlex 2200 Lyme Total Control Sets was used to evaluate reproducibility. Each of the panel members and a positive and negative control were tested in quadruplicate on 2 runs per day over 5 days at each of 3 sites (4 replicates x 2 runs x 5 days = 40 replicates per panel member per site = 120 total replicates for 3 sites). The data was analyzed for intra-assay and inter-assay reproducibility according to the CLSI guidance EP15-A3. Results are shown in the table below.
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| Lyme TotalPanel Member | N | Mean(AI) | Repeatability | Between Run | Between Day | Between Site | Total | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | |||
| Low Negative | 120 | 0.4 | 0.020 | 5.1% | 0.000 | 0.0% | 0.004 | 1.0% | 0.006 | 1.5% | 0.022 | 5.4% |
| High Negative | 120 | 0.7 | 0.035 | 5.1% | 0.013 | 1.8% | 0.023 | 3.3% | 0.032 | 4.5% | 0.055 | 7.8% |
| Low Positive | 120 | 1.1 | 0.040 | 3.6% | 0.016 | 1.5% | 0.027 | 2.4% | 0.075 | 6.7% | 0.091 | 8.1% |
| Mid Positive | 120 | 1.8 | 0.049 | 2.7% | 0.000 | 0.0% | 0.024 | 1.3% | 0.111 | 6.2% | 0.124 | 7.0% |
| High Positive | 120 | 4.7 | 0.108 | 2.3% | 0.116 | 2.5% | 0.121 | 2.6% | 0.131 | 2.8% | 0.239 | 5.1% |
BioPlex 2200 Lyme Total Reproducibility
Analytical Specificity: Analytical specificity of the BioPlex 2200 Lyme Total kit was evaluated against 836 samples from asymptomatic individuals from endemic regions. These samples were obtained from an asymptomatic population and excluded pre-screened blood donors. The results, expressed as % negatives, are presented in the following table.
BioPlex 2200 Lyme Total Analytical Specificity
| N | BioPlex 2200 Lyme Total | Commercial Lyme IgM/IgGImmunoassay | |
|---|---|---|---|
| AsymptomaticNon-Endemic | 416 | 96.4% | 95.4% |
| AsymptomaticEndemic | 420 | 96.4% | 94.8% |
Interfering Substances: An interfering substances study was conducted to evaluate the potential interference of specific endogenous and exogenous substances with the BioPlex 2200 Lyme Total kit according to CLSI EP07-A2 guideline. No interference was observed with any of the substances tested. The substances and the maximum levels tested are shown in the table below:
Interfering Substances
| Substance | Concentration |
|---|---|
| Hemoglobin | 1000 mg/dL |
| Bilirubin (unconjugated) | 20 mg/dL |
| Bilirubin (conjugated) | 30 mg/dL |
| Triglycerides | 3300 mg/dL |
| Total Protein | 12 g/dL |
| Cholesterol | 500 mg/dL |
| Ascorbic Acid | 6 mg/dL |
| EDTA | 800 mg/dL |
| Sodium Heparin | 8000 units/dL |
| Lithium Heparin | 8000 units/dL |
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Cross-Reactivity: A cross-reactivity study was performed to determine if samples from various disease states and other potentially cross-reacting agents interfere with test results when tested with the BioPlex 2200 Lyme Total kit. Samples known to be positive for each cross-reactant listed in the table below were evaluated with the BioPlex 2200 Lyme Total kit. All samples were pre-tested by a commercially available Lyme assay and only those that tested negative by the commercially available assay were further tested by the BioPlex 2200 Lyme Total kit. Percent negative agreement for each potential cross reactant is shown below.
| Negative BioPlex 2200 Results/Negative Commercial Lyme IgM/IgGImmunoassay Results | |||
|---|---|---|---|
| PotentialCross Reactant | N | %NegativeAgreement | |
| Anti-nuclear antibody (ANA) | 13/13 | 100% | |
| Babesiosis | 10/10 | 100% | |
| Chronic Fatigue Syndrome | 14/14 | 100% | |
| Cytomegalovirus | 13/13 | 100% | |
| Epstein-Barr Virus | 45/49 | 92% | |
| Ehrlichiosis | 10/11 | 91% | |
| HAMA | 13/13 | 100% | |
| Helicobacter pylori | 12/12 | 100% | |
| HIV | 13/13 | 100% | |
| Influenza virus | 14/15 | 93% | |
| Leptospirosis | 33/34 | 97% | |
| Multiple sclerosis | 14/14 | 100% | |
| Parvovirus B19 | 14/14 | 100% | |
| Periodontal | 34/35 | 97% | |
| Rheumatoid arthritis | 14/14 | 100% | |
| Rheumatoid Factor positive | 13/13 | 100% | |
| Rickettsial diseases | 16/16 | 100% | |
| Rubella | 14/14 | 100% | |
| Syphilis | 47/48 | 98% | |
| Systemic Lupus Erythematosus(SLE) | 15/15 | 100% | |
| Toxoplasmosis | 29/29 | 100% | |
| Varicella Zoster Virus | 14/14 | 100% |
Results of Cross-Reactivity Study
Matrix comparison: Matched serum and plasma (EDTA and heparin) samples drawn from the same donor were acquired through a vendor. A minimum of 40 sets of paired serum and plasma samples were prepared and values within the measurement range of the assay were analyzed. Samples were spiked with high titer Lyme samples to obtain concentrations that span the assay range. Samples were assayed in replicates of two with the second replicate run in reverse order. Mean plasma AI values were compared to matched mean serum AI values. Linear regression analysis was used to determine the presence of a matrix effect when compared to serum. The regression correlation parameters for
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slope, intercept and correlation coefficient (r) are shown below.
| Matrix Comparison | N | Slope(95% CI) | Intercept(95% CI) | Correlation(r) |
|---|---|---|---|---|
| K2-EDTAVS.Serum | 77 | 1.02(0.95 to 1.08) | 0.17(-0.09 to 0.44) | 0.960 |
| K3-EDTAVS.Serum | 74 | 1.01(0.93 to 1.08) | 0.14(-0.14 to 0.42) | 0.951 |
| Sodium HeparinVS.Serum | 77 | 0.97(0.91 to 1.02) | 0.09(-0.13 to 0.30) | 0.970 |
| Lithium HeparinVS.Serum | 78 | 0.97(0.92 to 1.01) | 0.05(-0.12 to 0.22) | 0.981 |
BioPlex 2200 Lyme Total Matrix Comparison
Clinical Studies
Method Comparison Study: Performance of the BioPlex 2200 Lyme Total kit was evaluated using prospectively collected serum samples from patients submitted for Lyme disease testing originating from various geographically distinct locations in the U.S. A total of 792 serum samples were evaluated at three (3) U.S. clinical testing sites. Equivocal (Eqv.) samples are added to positives for calculating agreement results since they will be tested by the second tier Western blot. Results are shown in the table below.
BioPlex 2200 Lyme Total Percent Agreement with Predicate Device - 1* Tier
| Commercial Lyme IgM/IgGImmunoassay | ||||||
|---|---|---|---|---|---|---|
| Pos | Eqv. | Neg | % Agreement | 95% CI | ||
| BioPlex 2200Lyme Total | Reactive | 65 | 0 | 30 | PPA = 70.1% (68/97) | 60.0 - 79.0% |
| Eqv. | 2 | 1 | 4 | |||
| Non-Reactive | 21 | 8 | 661 | NPA = 95.1% (661/695) | 93.2 - 96.6% | |
| Total | 88 | 9 | 695 |
PPA=positive percent agreement, NPA= negative percent agreement
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Second-Tier Testing: All reactive/positive and equivocal samples by the BioPlex 2200 Lyme Total kit and the commercially available predicate device were tested by FDAcleared IgG and IgM Western blot assays. Shown below are the second tier Western blot results (combined IgG and IgM) for samples that were reactive/ positive and equivocal in the prospective sample study.
| Tier 1+ or ± | WB + | WB - | |
|---|---|---|---|
| Commercial LymeIgM/IgGImmunoassay | 97 | 47 | 50 |
| BioPlex 2200Lyme Total | 102 | 49 | 53 |
| Commercial LymeIgM/IgGImmunoassay +BioPlex 2200Lyme Total | 68 | 46 | 22 |
| Second Tier Testing and PPA Analysis | |||||
|---|---|---|---|---|---|
| -- | -- | -- | -- | -- | -------------------------------------- |
| 1st Tier PPA(95% CI) | 70.1%(60.0-79.0%) | 68/97 |
|---|---|---|
| 2nd Tier PPA(95% CI) | 97.9%(88.7-99.9%) | 46/47 |
Sensitivity Study: Sensitivity of the BioPlex 2200 Lyme Total kit was evaluated using 105 clinically characterized samples from an archived collection containing early, convalescent and late phases of Lyme disease. The table below shows the sensitivity of the BioPlex 2200 Lyme Total kit for each of the disease phases along with a commercial Lyme IgM/IgG immunoassay.
| BioPlex 2200 Lyme Sensitivity with respect to Disease Stage | ||
|---|---|---|
| Clinical Stage | N | BioPlex 2200 Lyme Total | Sensitivity95% CI | Commercial Lyme IgM/IgG Immunoassay | Sensitivity95% CI | ||||
|---|---|---|---|---|---|---|---|---|---|
| Reactive | Equivocal | Non-Reactive | Positive | Equivocal | Negative | ||||
| Acute(< 3 months) | 72 | 50 | 0 | 22 | 69.4%(50/72)57.5 — 79.8% | 42 | 0 | 30 | 58.3%(42/72)46.1 — 69.9% |
| Convalescent(< 12 months) | 26 | 16 | 0 | 10 | 61.5%(16/26)40.6 — 79.8% | 17 | 1 | 8 | 69.2%(18/26)48.2 — 85.7% |
| Late(> 12 months) | 7 | 6 | 0 | 1 | 85.7%(6/7)42.1 — 99.6% | 5 | 0 | 2 | 71.4%(5/7)29.0 — 96.3% |
| Total | 105 | 72 | 0 | 33 | 68.6%(72/105)58.8 — 77.3% | 64 | 1 | 40 | 61.9%(65/105)51.9 — 71.2% |
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CDC Panel: A panel of 280 positive and negative specimens from the Centers for Disease Control and Prevention (CDC) was tested for the presence of Lyme antibodies using the BioPlex 2200 Lyme Total kit. The results are presented as a means to convey further information on the performance of the BioPlex 2200 Lyme Total kit with a masked, characterized serum panel. This does not imply an endorsement of the BioPlex 2200 Lyme Total kit by the CDC. Results are summarized in the table below.
| BioPlex 2200 Lyme Total | |||||
|---|---|---|---|---|---|
| (N=280) | N | Reactive | Equivocal | Non-Reactive | % Agreement withClinical Diagnosis |
| Acute | 39 | 33 | 0 | 6 | 84.6% |
| Convalescent | 31 | 29 | 0 | 2 | 93.5% |
| Late | 20 | 20 | 0 | 0 | 100% |
| Look-alike Diseases | 90 | 1 | 2 | 87 | 96.7% |
| Healthy Controls | 100 | 1 | 2 | 97 | 97.0% |
BioPlex 2200 Lyme Total Testing of CDC Reference Sera
The CDC panel was further evaluated against the BioPlex 2200 Lyme Total kit and a commercially available predicate device. The results are summarized in the table below.
| BioPlex 2200 Lyme Total Comparative Testing of CDC Reference Sera | |||
|---|---|---|---|
| Sample Category(N=280) | N | BioPlex 2200 Lyme Total | Commercial Lyme IgM/IgGImmunoassay | |||||||
|---|---|---|---|---|---|---|---|---|---|---|
| R(+) | Eqv | NR(-) | %Agreement | Pos(+) | Eqv | Neg(-) | %Agreement | |||
| Early EM | Acute | 30 | 24 | 0 | 6 | 80.0% | 19 | 0 | 11 | 63.3% |
| Convalescent | 30 | 28 | 0 | 2 | 93.3% | 27 | 0 | 3 | 90.0% | |
| Cardiac Lyme | Acute | 3 | 3 | 0 | 0 | 100% | 3 | 0 | 0 | 100% |
| NeurologicalLyme | Acute | 6 | 6 | 0 | 0 | 100% | 6 | 0 | 0 | 100% |
| Convalescent | 1 | 1 | 0 | 0 | 100% | 1 | 0 | 0 | 100% | |
| LymeArthritis/Neuro. | Late | 20 | 20 | 0 | 0 | 100% | 20 | 0 | 0 | 100% |
| Fibromyalgia | Look-alikeDiseases | 15 | 0 | 1 | 14 | 93.3% | 0 | 0 | 15 | 100% |
| Rheumatoidarthritis | 15 | 0 | 0 | 15 | 100% | 0 | 0 | 15 | 100% |
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| Sample Category(N=280) | N | BioPlex 2200 Lyme Total | Commercial Lyme IgM/IgGImmunoassay | |||||||
|---|---|---|---|---|---|---|---|---|---|---|
| R(+) | Eqv | NR(-) | %Agreement | Pos(+) | Eqv | Neg(-) | %Agreement | |||
| Multiplesclerosis | 15 | 0 | 0 | 15 | 100% | 0 | 0 | 15 | 100% | |
| Mononucleosis | 15 | 0 | 2 | 13 | 86.7% | 2 | 0 | 13 | 86.7% | |
| Syphilis | 15 | 0 | 0 | 15 | 100% | 1 | 0 | 14 | 93.3% | |
| Periodontitis | 15 | 0 | 0 | 15 | 100% | 0 | 0 | 15 | 100% | |
| EndemicNegativeControls | HealthyNormals | 50 | 0 | 2 | 48 | 96.0% | 0 | 0 | 50 | 100% |
| Non-EndemicNegativeControls | 50 | 1 | 0 | 49 | 98.0% | 1 | 0 | 49 | 98.0% |
R= Reactive, NR= Nonreactive
Conclusion: The submitted information in this 510(k) Premarket Notification will support a determination of substantial equivalence for BioPlex 2200 Lyme Total. The clinical and non-clinical tests demonstrate that this device is safe and effective and performs as well as the legally marketed predicate device.
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Indications for Use
510(k) Number (if known)
Device Name
Indications for Use (Describe)
Type of Use (Select one or both, as applicable)Prescription Use (Part 21 CFR 801 Subpart D)
| Over-The-Counter Use (21 CFR 801 Subpart C)
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§ 866.3830
Treponema pallidum treponemal test reagents.(a)
Identification. Treponema pallidum treponemal test reagents are devices that consist of the antigens, antisera and all control reagents (standardized reagents with which test results are compared) which are derived from treponemal sources and that are used in the fluorescent treponemal antibody absorption test (FTA-ABS), theTreponema pallidum immobilization test (T.P.I.), and other treponemal tests used to identify antibodies toTreponema pallidum directly from infecting treponemal organisms in serum. The identification aids in the diagnosis of syphilis caused by bacteria belonging to the genusTreponema and provides epidemiological information on syphilis.(b)
Classification. Class II (performance standards).