(93 days)
The Borrelia burgdorferi IgG/IgM ELISA kit is an Enzyme-Linked Immunosorbent Assay (ELISA) for the qualitative presumptive detection of total (IgG/IgM) antibodies to Borrelia burgdorferi in human serum. This ELISA should only be used for patients with signs and patients with symptoms that are consistent with Lyme disease. Equivocal or positive results should be supplemented by testing with a standardized Western blot procedure. Positive supplemental results are supportive evidence of exposure to B. burgdorferi and can be used to support a clinical diagnosis of Lyme disease.
The Borrelia burgdorferi IgG/IgM ELISA test is an enzyme linked immunosorbent assay to detect IgGM antibodies to Borrelia burgdorferi. Purified Borrelia burgdorferi antigen is attached to a solid phase microtiter well. Diluted test sera is added to each well. If the antibodies are present that recognize the antigen, thev will bind to the antigen in the well. After incubation the wells are washed to remove unbound antibodv. An enzyme labeled anti-human IgG/IgM is added to each well. If antibody is present it will bind to the antibody attached to the antigen on the well. After incubation the wells are washed to remove unbound conjugate. A substrate solution is added to each well. If enzyme is present the substrate will undergo a color change. After an incubation period the reaction is stopped and the color intensity is measured photometrically, producing an indirect measurement of specific antibody in the patient specimen.
Here's a breakdown of the acceptance criteria and study information for the Borrelia burgdorferi IgG/IgM ELISA Test Kit, based on the provided text:
1. Table of Acceptance Criteria and Reported Device Performance
The document doesn't explicitly state formal "acceptance criteria" with specific numerical thresholds for all performance metrics. However, we can infer the desired performance and report the observed results.
| Acceptance Criterion (Inferred) | Reported Device Performance | Section from Document |
|---|---|---|
| Clinical Sensitivity (agreement with clinical diagnosis for Lyme disease) | 71% (30/42) | Wampole B. burgdorferi IgG/IgM ELISA Result |
| Clinical Specificity (agreement with negative diagnosis for normals) | 100% (5/5) | Table 1 The CDC Lyme Disease Serum Panel Stratified by Time After Onset. |
| Agreement with Predicate Device (Lyme STAT) post 2-step testing | Wampole: 4% (1.0%-6.9%) (7/176) Lyme Stat: 2.8% (0.3%-5.3%) (5/176) | Study 2: Table 2 |
| Precision (Intersite CV) | Generally <15% CV (with two outliers at 40.57% and 68.93%) | Table 3 and associated text |
| False Positives in Precision Study | 0 (no positive results for negative sera) | Table 3 and associated text (last paragraph) |
| False Negatives in Precision Study | 0 (no negative results for positive sera) | Table 3 and associated text (last paragraph) |
| Cross-Reactivity | Low (1 positive for Bilirubinemic, 1 positive for Cytomegalovirus Antibody +) | Table 4 Cross Reactivity |
2. Sample Size Used for the Test Set and Data Provenance
- Test Set 1 (CDC Lyme Disease Serum Panel):
- Sample Size: 47 sera (5 normals, 42 Lyme disease patients).
- Data Provenance: Serum panel obtained from the CDC. The text mentions "masked, characterized serum panel." This is a retrospective collection. The country of origin is not explicitly stated but implied to be the US given the CDC involvement.
- Test Set 2 (Comparative Study with Predicate):
- Sample Size: 176 fresh sera.
- Data Provenance: "fresh sera from patients of various ages and genders that were submitted to a large clinical lab for B. burgdorferi antibody testing." This implies a prospective collection from a clinical lab, likely in the US.
- Test Set 3 (Precision Study):
- Sample Size: 7 sera (tested 10 times each on 3 plates at 3 sites = 630 determinations) + 3 sera (tested 10 times each on 3 plates at 2 sites = 180 determinations) + 3 controls (HPC, LPC, NC, 15 determinations each) + 1 calibrator (CAL, 45 determinations). Total determinations mentioned as "810 determinations". The actual number of unique sera is 10.
- Data Provenance: Not explicitly stated, but clinical lab samples are implied for the sera, likely in the US.
- Test Set 4 (Cross-Reactivity Study):
- Sample Size: 58 samples across various potentially cross-reactive conditions (e.g., 5 lipemic, 5 bilirubinemic, 10 RPR+ etc.).
- Data Provenance: Not explicitly stated, but likely clinical samples. No country of origin mentioned.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of Those Experts
- Test Set 1 (CDC Panel): The ground truth is described as "Clinical Diagnosis" for the Lyme disease patients and "normals" for the controls. The document doesn't specify the number or qualifications of experts involved in establishing these clinical diagnoses for the CDC panel.
- Test Set 2 (Comparative Study): The ground truth for positive/equivocal Wampole/Lyme Stat results was established by Western Blot testing (Mardx Diagnostics Western Blot on both IgG and IgM). This is a laboratory-based method, not a direct expert consensus on the diagnosis itself. The interpretation of the Western Blot, however, would typically be done by trained laboratory personnel.
- Test Set 3 & 4: Ground truth pertains to the known characteristics of the samples (e.g., known positive/negative for precision, known condition for cross-reactivity). No human experts were involved in establishing "ground truth" for the device's output to be compared against.
4. Adjudication Method for the Test Set
- Test Set 1 (CDC Panel): Not explicitly stated. The "Clinical Diagnosis" is the reference, but how that diagnosis was made or adjudicated for the panel is not detailed.
- Test Set 2 (Comparative Study): For samples that were "positive or equivocal" by either the Wampole or Lyme Stat ELISA, Western Blot (WB) was used as the confirmatory test. This serves an adjudicative role by providing a higher-tier diagnostic confirmation. There is no mention of human expert adjudication beyond the Western Blot result itself.
- Test Set 3 & 4: Not applicable, as these studies are performance characterizations against known sample properties.
5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was Done
No, a Multi-Reader Multi-Case (MRMC) comparative effectiveness study was not done. The studies presented focus on the standalone performance of the device or its comparison to another device and Western blot as a gold standard, not on how human readers' performance might improve with or without AI assistance. The device is a diagnostic assay, not an AI-powered image analysis tool.
6. If a Standalone (i.e., algorithm only without human-in-the-loop performance) was Done
Yes, the studies are primarily standalone performance evaluations of the ELISA test kit. The ELISA is a laboratory assay that produces a quantitative (optical density) result, which is then interpreted qualitatively (positive/negative/equivocal) based on predefined cut-offs. There is no "human-in-the-loop" interaction in the sense of an AI system providing assistance. The interpretation of the ELISA results for patient management would involve human clinicians, but the performance characteristics detailed here are for the assay itself.
7. The Type of Ground Truth Used
- Test Set 1 (CDC Panel): Clinical Diagnosis. This implies a physician's assessment based on symptoms, signs, and potentially other diagnostic tests.
- Test Set 2 (Comparative Study): Western Blot (laboratory-based confirmatory test). For comparison against the ELISA, the Western Blot was used as a more definitive ground truth for antibody presence.
- Test Set 3 (Precision Study): Internal controls or characterized sera with known expected values/positivity.
- Test Set 4 (Cross-Reactivity Study): Samples characterized by the presence of specific interfering substances or conditions (e.g., "Lipemic (+++)", "RPR +", etc.).
8. The Sample Size for the Training Set
The document does not mention or describe a "training set." This device is a traditional ELISA test kit, not a machine learning or AI algorithm that requires a training phase. Its performance is based on the biochemical reaction and pre-defined cut-off values, not on a learned model.
9. How the Ground Truth for the Training Set Was Established
Not applicable, as there is no training set for this type of device.
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Summary of Safety and Effectiveness Information Borrelia burgdorferi IgG/IgM ELISA Test Kit
I. Immuno Probe Inc. 1306 Bailes Lane, Suite F Frederick, Maryland 21701 Contact person: William Boteler Telephone: 301-695-7920 Date of preparation: March 4, 1997
MAR 26 1997
II. Description of Device
The Borrelia burgdorferi IgG/IgM ELISA kit is an Enzyme-Linked Immunosorbent Assay (ELISA) for the qualitative presumptive detection of total (IgG/IgM) antibodies to Borrelia burgdorferi in human serum. This ELISA should only be used for patients with signs and patients with symptoms that are consistent with Lyme disease. Equivocal or positive results should be supplemented by testing with a standardized Western blot procedure. Positive supplemental results are supportive evidence of exposure to B. burgdorferi and can be used to support a clinical diagnosis of Lyme disease.
The Borrelia burgdorferi IgG/IgM ELISA test is an enzyme linked immunosorbent assay to detect IgGM antibodies to Borrelia burgdorferi. Purified Borrelia burgdorferi antigen is attached to a solid phase microtiter well. Diluted test sera is added to each well. If the antibodies are present that recognize the antigen, thev will bind to the antigen in the well. After incubation the wells are washed to remove unbound antibodv. An enzyme labeled anti-human IgG/IgM is added to each well. If antibody is present it will bind to the antibody attached to the antigen on the well. After incubation the wells are washed to remove unbound conjugate. A substrate solution is added to each well. If enzyme is present the substrate will undergo a color change. After an incubation period the reaction is stopped and the color intensity is measured photometrically, producing an indirect measurement of specific antibody in the patient specimen.
III. Predicate Device
The Borrelia burgdorferi IgG/IgM ELISA test is substantially equivalent to BioWhittaker's Lyme STAT test. Equivalence is demonstrated by the following comparative results:
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Performance Characteristics
Table 1 The CDC Lyme Disease Serum Panel Stratified by Time After Onset.
The following information is from a serum panel obtained from the CDC and tested by Wampole. The 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.
| Time FromOnset | + | Equivocal | - | Total | % Agreementwith ClinicalDiagnosis |
|---|---|---|---|---|---|
| normals | 0 | 0 | 5 | 5 | 100% |
| <1 month | 2 | 0 | 3 | 5 | 40.0% |
| 1-2 months | 6 | 2 | 2 | 10 | 80.0% |
| 3-12 months | 9 | 3 | 7 | 19 | 63.3% |
| > 1yr | 8 | 0 | 0 | 8 | 100.0% |
| Total | 25 | 5 | 17 | 47 |
Wampole B. burgdorferi IgG/IgM ELISA Result
Because the equivocals would have been tested bv immunoblotting in a 2-step test system, they were considered 1-step positive for purposes of calculating % agreement. The Wampole Borrelia burgdorferi IgGflgM ELISA demonstrated 71% (30/42) agreement (sensitivity) with clinical diagnosis of Lyme disease.
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Study 2: 176 fresh sera from patients of various ages and genders that were submitted to a large clinical lab for B. burgdorferi antibody testing were tested on the Wampole B. burgdorferi IgG/IgM ELISA and Lyme Stat. Any serum found positive or equivocal was tested by Mardx Diagnostics Western Blot on both IgG and IgM.
Table 2: Western blot results
| Western Blot | |||
|---|---|---|---|
| + | - | ||
| Wampole | + | 6 | 3 |
| eq. | 1 | 6 | |
| Lyme Stat | + | 5 | 3 |
| eq. | 0 | 5 |
| Type of Result | Wampole (95%CI) | Lyme Stat (95%CI) |
|---|---|---|
| 1-step (ELISA) pos. or eq. | 9.1% (4.8%-13.4%)(16/176) | 7.4% (3.4%-11.3%)(13/176) |
| 1-step pos. or eq. &2-step (WB) pos. | 4% (1.0%-6.9%)(7/176) | 2.8% (0.3%-5.3%)(5/176) |
| 2-step pos. among1-step pos. or eq. | 44% (18.9%-68.6%)(7/16) | 39% (11.5%-65.4%(5/13) |
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- Precision. Seven sera were assayed ten times each on three different plates at three different sites. An additional three serum were assayed ten times each on three different plates at two different sites. The intersite precision is shown in Table 3. With appropriate technique the user should obtain precision of <15% CV.
| Table 3 | ||
|---|---|---|
| Borrelia burgdorferi IgG/IgM ELISA Inter Assay Precision Between Sites |
| Inter-Assay (n=90) | ||||
|---|---|---|---|---|
| # | X | SD | CV | n |
| 1. | 1.18 | 0.113 | 9.58% | 90 |
| 2. | 2.26 | 0.226 | 10.02% | 90 |
| 3. | 3.85 | 0.278 | 7.21% | 90 |
| 4. | 5.36 | 0.402 | 7.50% | 90 |
| 5. | 2.04 | 0.186 | 9.11% | 90 |
| 6. | 0.33 | 0.132 | 40.57% | 90 |
| 7. | 0.34 | 0.234 | 68.93% | 90 |
| 8. | 1.47 | 0.152 | 10.34% | 60 |
| 9. | 1.47 | 0.104 | 7.10% | 60 |
| 10. | 1.65 | 0.131 | 7.95% | 60 |
| HPC | 5.31 | 0.314 | 5.91% | 15 |
| LPC | 2.70 | 0.107 | 3.96% | 15 |
| NC | 0.20 | 0.043 | 21.97% | 15 |
| CAL | 3.34 | 0.103 | 3.08% | 45 |
A total of 810 determinations were made at the three sites. In all 810 determinations there was not a cas a positive result for a negative serum or a negative result for a positive serum.
X = Mean ISR SD = standard deviation CV = coefficient of variation = SD/X x 100 The methods in NCCLS EP5 were utilized for precision parameters.
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- Cross-Reactivity. The following potentially cross-reactive sera were run on the Borrelia burgdorferi IgG/IgM ELISA assay to assess cross-reactivity with the assay: lipemic, bilirubinemic, RPR +, dsDNA +, RF +, EBV +, CMV +, RMS +, elevated ESR, and CRP. The data in Table 4 illustrates the amount of reactivity with the sera.
TABLE 4
Cross Reactivity
| Laboratory result (Titer) | # of samples | # of positives |
|---|---|---|
| Lipemic (+++) | 5 | 0 |
| Bilirubinemic (1.9-16.9) | 5 | 1 |
| RPR + (1:2 - 1:64) | 10 | 0 |
| Rheumatoid Factor + (1:40-1:320) | 3 | 0 |
| Epstein Barr Virus Antibody + (1:40-1:2560) | 7 | 0 |
| Cytomegalovirus Antibody + (O.D. 0.718-2.308) | 6 | 1 |
| Rocky Mt Spotted Fever Antibody + (1:256-1:16,384) | 4 | 0 |
| CRP + (2.79-8.61 mg/dl) | 5 | 0 |
| Elevated ESR (40-115) | 10 | 0 |
| dsDNA + (52.3-1072 IU) | 16 | 0 |
§ 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).