The Borrelia burgdorferi IgM ELISA kit is an Enzyme-Linked Immunosorbent Assay (ELISA) for the qualitative presumptive detection of IgM antibodies to Borrelia burgdorferi in human serum. This ELISA should only be used for patients with signs and symptoms that are consistent with Lyme disease. Equivocal or positive results must 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.

Device Description

The Borrelia burgdorferi IgM ELISA test is an enzyme linked immunosorbent assay to detect IgM antibodies to Borrelia burgdorferi. Purified Borrelia burgdorferi antigen is attached to a solid phase microtiter well. Pretreated test sera is added to each well. If the antibodies are present that recognize the antigen, they will bind to the antigen in the well. After incubation the wells are washed to remove unbound antibody. An enzyme labeled anti-human 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.

AI/ML Overview

Here's an analysis of the provided information regarding the Borrelia burgdorferi IgM ELISA Test Kit, structured according to your request.

Device Name: Borrelia burgdorferi IgM ELISA Test Kit

1. Table of Acceptance Criteria and Reported Device Performance

It's important to note that the provided documents do not explicitly state "acceptance criteria" as clear pass/fail thresholds. Instead, they present performance characteristics from various studies. Based on the provided data, I have inferred what could be considered the reported performance of the device in these studies.

Acceptance Criteria (Inferred)	Reported Device Performance (Wampole Borrelia burgdorferi IgM ELISA)
Sensitivity (Agreement with Clinical Diagnosis) for Lyme Disease (CDC Panel)	47.6% (20/42) when equivocals are considered 1-step positive.
Agreement with Clinical Diagnosis for Normals (CDC Panel)	100% (5/5)
Agreement with Clinical Diagnosis for <1 month post-onset (CDC Panel)	80.0% (4/5)
Agreement with Clinical Diagnosis for 1-2 months post-onset (CDC Panel)	60.0% (6/10)
Agreement with Clinical Diagnosis for 3-12 months post-onset (CDC Panel)	47.4% (9/19)
Agreement with Clinical Diagnosis for >1 year post-onset (CDC Panel)	12.5% (1/8)
2-step positive rate (ELISA pos/eq & Western Blot pos) among total tested in a real-world setting	3.41% (6/176)
2-step positive rate (ELISA pos/eq & Western Blot pos) among 1-step pos or eq in a real-world setting	31.6% (6/19)
Inter-site precision (CV)	Generally < 20% CV for most samples (e.g., 8.96%, 9.64%, 11.2%, 15.82%, 13.96%, 17.18%). Exceptions: Sample #4 (68.0% CV) and NC (64.00% CV). HP (7.04%), Cal (4.70%), LP (5.11%). "With appropriate technique the user should obtain precision of <20% CV." indicating this as a target.
Cross-reactivity	0 positives observed for all tested potentially cross-reactive sera (Lipemic, Bilirubinemic, RPR+, RF+, EBV+, CMV+, RMS+, CRP+, Elevated ESR, dsDNA+).
No positive result for a negative specimen and no negative result for a positive specimen in precision study (false positives/negatives)	Not a single case reported across 492 determinations.

2. Sample size used for the test set and the data provenance

Study 1 (CDC Lyme Disease Serum Panel):
- Sample Size: 47 serum samples.
- Data Provenance: Obtained from the CDC, characterized (masked), origin not specified, but typically such panels would represent a diverse set of patients. This is likely a retrospective panel.
Study 2 (Clinical Lab Sera):
- Sample Size: 176 fresh sera.
- Data Provenance: Patients from a "large clinical lab" submitted for B. burgdorferi antibody testing. The term "fresh sera" suggests a more prospective collection in a clinical setting at that time. The country of origin is not specified but is implied to be within the US given the submission to the FDA.
Study 3 (Precision Study):
- Sample Size: 7 sera, each assayed 10 times, on 3 different assays, at 2 different sites. This implies 7 * 10 * 3 * 2 = 420 individual tests for the 7 sera. Additionally, HP, LP, NC had n=12 (per site), and Cal had n=36 (per site). Total of 492 determinations.
- Data Provenance: Not specified, but likely laboratory-prepared samples / controls.
Study 4 (Cross-Reactivity):
- Sample Size: 53 samples (5 Lipemic, 5 Bilirubinemic, 10 RPR+, 8 RF+, 7 EBV+, 6 CMV+, 4 RMS+, 5 CRP+, 10 Elevated ESR, 16 dsDNA+).
- Data Provenance: Not specified, but these would be known positive samples for the interfering substances. Likely retrospective collection from various sources.

3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts

Study 1 (CDC Lyme Disease Panel): The ground truth is referred to as "Clinical Diagnosis." The information doesn't specify the number or qualifications of experts involved in establishing these clinical diagnoses for the CDC panel. It is implied that the panel had a pre-established characterization based on clinical criteria and likely expert review.
Study 2 (Clinical Lab Sera):
- The ground truth for this study was established using Western Blot (Mardx Diagnostics Western Blot on both IgG and IgM) for any serum found positive or equivocal by ELISA.
- The document does not specify the number of experts, if any, involved in interpreting these Western Blots or in establishing the initial clinical diagnoses for these 176 fresh sera. Western blot interpretation itself often relies on established guidelines, which may implicitly incorporate expert consensus.

4. Adjudication method for the test set

The document does not explicitly describe an adjudication method (like 2+1 or 3+1 consensus) for establishing the ground truth definitions in any of the studies.
For Study 1, the "Clinical Diagnosis" assigned to the CDC panel samples serves as the ground truth.
For Study 2, Western Blot results were used as a confirmatory "ground truth" for positive/equivocal ELISA samples. This is a technical adjudication rather than a human consensus adjudication on images.

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

No, a multi-reader multi-case (MRMC) comparative effectiveness study was not performed.
This device is an ELISA lab test kit, not an AI-assisted diagnostic imaging or interpretation tool. Therefore, the concept of "human readers improve with AI vs without AI assistance" does not apply here.

6. If a standalone (i.e., algorithm only without human-in-the-loop performance) was done

Yes, the performance presented for the Wampole Borrelia burgdorferi IgM ELISA is a standalone performance of the test kit itself. It measures the direct output of the assay ("positive", "equivocal", "negative") based on antibody detection.
The human role is in performing the lab procedure, reading the photometric results, and interpreting those results based on the kit's cut-offs, but the core "decision" of the assay (antibody presence) is algorithmic based on chemical reactions and optical density.

7. The type of ground truth used (expert consensus, pathology, outcomes data, etc.)

Study 1 (CDC Panel): "Clinical Diagnosis" stratified by time after onset. This implicitly involves expert clinical judgment and the patient's presentation/history.
Study 2 (Clinical Lab Sera): Western blot results (IgM and IgG) for samples that were positive or equivocal by ELISA. This is a higher-tier laboratory confirmatory test.
It's important to note that the ELISA kit itself is for "qualitative presumptive detection" and "equivocal or positive results must be supplemented by testing with a standardized Western blot procedure," indicating the regulatory recognition of Western Blot as a more definitive confirmatory step in the diagnostic pathway.

8. The sample size for the training set

The document does not provide information about a separate "training set" for the device. This is typical for traditional ELISA kits, which are based on biochemical principles and established reagent formulations rather than machine learning algorithms that require explicit training data. The development and optimization of the assay would have involved various internal sample testing, but this wouldn't be referred to as a "training set" in the context of AI.

9. How the ground truth for the training set was established

As there's no mention of a "training set" in the context of the device being an AI/ML algorithm, this question is not applicable. The development of the ELISA kit would have relied on known positive and negative controls, and extensive R&D, rather than a training set with established ground truth in the AI sense.

Summary

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K966129

Summary of Safety and Effectiveness Information Borrelia burgdorferi IgM ELISA Test Kit

MAR 26 1997

I. Immuno Probe Inc. 1306 Bailes Lane, Suite F Frederick, Maryland 21701 Contact person: William Boteler Telephone: 301-695-7920 Date of preparation: March 17, 1997

II. Description of Device

The Borrelia burgdorferi IgM ELISA kit is an Enzyme-Linked Immunosorbent Assay (ELISA) for the qualitative presumptive detection of IgM antibodies to Borrelia burgdorferi in This ELISA should only be used for patients with signs and symptoms that are human serum. consistent with Lyme disease. Equivocal or positive results must 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.

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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	% Agreement withClinical Diagnosis
normals	0	0	5	5	100%
<1 month	3	1	1	5	80.0%
1-2 months	6	0	4	10	60.0%
3-12 months	7	2	10	19	47.4%
> 1yr	1	0	7	8	12.5%
Total	17	3	27	47

Wampole B. burgdorferi IgM ELISA Result

Because the equivocals would have been tested by immunoblotting in a 2-step test system, they were considered 1-step positive for purposes of calculating % agreement. The Wampole Borrelia burgdorferi IgM ELISA demonstrated 47.6% (20/42) 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 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	10
	eq.	0	3
Lyme Stat	+	6	9

eq. 0 3

Type of Result	Wampole	(95%CI)	Lyme Stat	(95%CI)
1-step (ELISA) pos. or eq.	10.8%(19/176)	(6.1-15.5%)	10.2%(18/176)	(5.7-14.8%)
1-step pos. or eq. &	3.41%	(0.7-6.1%)	3.41%	(0.7-6.1%)
2-step (WB) pos.	3.41%(6/176)		3.41%(6/176)
2-step pos. among1-step pos. or eq.	31.6%(6/19)	(10.3-52.9%)	33.3%(6/18)	(11.1-55.6%)

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Precision. Seven sera were assayed ten times each on three different assays at two different sites. The intersite precision is shown in Table 3. With appropriate technique the user should obtain precision of <20% CV.

Table 3 Comparison of Borrelia burgdorferi IgM ELISA Intra & Inter Assay Precision Between Sites

#	Inter-Assay (n=60)
	X	SD	CV
1	3.74	0.335	8.96%
2	1.95	0.188	9.64%
3	0.94	0.105	11.2%
4	0.05	0.034	68.0%
5	0.98	0.155	15.82%
6	1.34	0.187	13.96%
7	1.24	0.213	17.18%
HP *	4.07	0.287	7.04%
Cal **	2.22	0.104	4.70%
LP *	2.21	0.113	5.11%
NC *	0.05	0.031	64.00%

For HP, LP, and NC n = 12

** For Cal n = 36

A total of 492 determinations were made at the two sites. In all of the determinations there was not a case of a positive result for a negative specimen or a negative result for a positive specimen. Equivocal sample #3 was negative 23 times and positive 19 times. Equivocal sample #5 was negaitve 19 times and positive 3 times.

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 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	0
RPR + (1:2 - 1:64)	10	0
Rheumatoid Factor + (1:40-1:320)	8	0
Epstein Barr Virus Antibody + (1:40-1:2560)	7	0
Cytomegalovirus Antibody + (O.D. 0.718-2.308)	6	0
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

Regulation Number and Section

§ 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).