(391 days)
The ImmunoProbe anti-EBNA-1 IgG assay may be used in conjunction with other Epstein-Barr serologies (VCA IgG, VCA IgM, EA (R&D), and heterophile) as an aid in the diagnosis of infectious mononucleosis. For In Vitro Diagnostic Use Only.
The Epstein-Barr Virus Nuclear Antigen 1 (EBNA-1) IgG kit is an Enzyme-Linked Immunosorbent Assays (ELISA) for the semi-quantitative determination of IgG antibodies in human serum to EBNA-1 antigen. The EBNA-1 IgG EIA test is an enzyme linked immunosorbent assay to detect IgG antibodies to EBNA-1. Purified recombinant EBNA-1 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, they will bind to the antigen in the well. After incubation the wells are washed to remove unbound antibody. An enzyme labeled anti-human IgG 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.
This document describes the performance characteristics of the Immuno Probe EBNA-1 IgG EIA Test Kit (K951549) and presents studies to demonstrate its safety and effectiveness.
1. Table of Acceptance Criteria and Reported Device Performance
The acceptance criteria for the EBNA-1 IgG EIA Test Kit are not explicitly stated as distinct, pre-defined thresholds in the provided text. Instead, the document presents performance characteristics and demonstrates that the device's results are in line with a predicate device and expected clinical serological profiles for EBNA-1 IgG. Based on the "Performance Characteristics" section, we can infer the following performance metrics and observed results:
| Performance Metric | Acceptance Criteria (Inferred) | Reported Device Performance |
|---|---|---|
| Relative Sensitivity (vs. predicate) | High sensitivity (>90%) | 97.8% (95% CI: 96.1% - 99.6%) |
| Relative Specificity (vs. predicate) | High specificity (>90%) | 100% (95% CI: 96.0% - 100%) |
| Relative Agreement (vs. predicate) | High agreement (>90%) | 98.3% (95% CI: 96.9% - 99.7%) |
| Inter-site Precision (CV%) | Low variability (0.95 typically) | 0.97 - 0.98 |
| Cross-Reactivity | No cross-reactivity | No cross-reactivity observed with HSV I & II, CMV, and VZV |
| Sensitivity for 4-fold antibody rise | 100% detection | 100% for instances where acute sera had a value 90%) |
| Specificity (vs. serum characterization) | High specificity (>90%) | Inferred from agreement in acute/seronegative groups |
| Accuracy/Agreement (vs. serum characterization) | High agreement (>90%) | 94.7% (95% CI: 91.0%-98.3%) |
2. Sample size used for the test set and the data provenance
- Relative sensitivity and specificity (vs. predicate):
- Sample Size: 350 total specimens (276 positive, 74 negative). 8 equivocal results were excluded.
- Data Provenance: The study was conducted at three different sites: two R&D laboratories at commercial companies (Maryland and New York) and one large commercial lab (Pennsylvania). The data is retrospective, as it compares the new device's performance against an "alternate commercially available EIA assay" implying existing specimens.
- Precision:
- Sample Size: For each of the four sera (High Positive, Mid Positive, Low Positive, Negative) and two controls (Calibrator, High Positive Control), samples were tested 3 times, twice a day for 20 days at 3 sites.
- This totals to 3 * 2 * 20 * 3 = 360 individual assay points for each serum type across all sites (though the table states n=360 for "Inter Site Precision", implying this is the total number of data points aggregated for the CV calculation, not necessarily 360 distinct patient samples).
- Calibrator: n=240
- High Positive Control: n=120
- Data Provenance: Three different sites (locations not specified beyond "different sites"). Likely prospective as these were controlled runs of known sera/controls.
- Sample Size: For each of the four sera (High Positive, Mid Positive, Low Positive, Negative) and two controls (Calibrator, High Positive Control), samples were tested 3 times, twice a day for 20 days at 3 sites.
- Linearity:
- Sample Size: 5 different sera were serially diluted and tested.
- Data Provenance: Not specified, but likely laboratory-generated data as it involves controlled serial dilutions.
- Cross-Reactivity:
- Sample Size: 5 sera known to contain antibodies to HSV I & II, CMV, and VZV.
- Data Provenance: Not specified, but likely laboratory-generated data from specific known-positive samples.
- Evaluation of paired sera:
- Sample Size: 20 high positive sera were serially diluted to create 68 paired samples with a four-fold dilution.
- Data Provenance: Not specified, but likely laboratory-generated from existing high-positive samples.
- Sensitivity and Specificity Based on Serum Characterization:
- Sample Size: 150 specimens (95 seropositive, 24 acute, 31 seronegative). 8 equivocal results were excluded.
- Data Provenance: The serum was from "the first study site" which implies one of the sites mentioned in the relative sensitivity/specificity study. The characterization (seronegative, acute, seropositive) suggests these were clinical samples. The study is likely retrospective, using samples with pre-established serological profiles.
3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts
The document does not explicitly state the number or qualifications of experts used to establish the ground truth.
- For Relative Sensitivity/Specificity: The ground truth for the comparison was an "alternate commercially available EIA assay." It is implied that the results from this predicate device are considered the truth for this comparison. The "experts" would implicitly be the developers and users of this predicate device, whose qualifications are not detailed.
- For Sensitivity/Specificity Based on Serum Characterization: The ground truth was established by classifying sera as "seronegative," "acute," or "seropositive" based on "other Epstein-Barr serologies (VCA IgG, VCA IgM)." This implies a determination made by laboratory personnel or serologists who follow established diagnostic protocols. No specific number or qualifications are provided.
4. Adjudication method for the test set
The document does not describe an adjudication method involving multiple readers for the test sets. The results appear to be based on direct laboratory measurements and comparisons to a predicate device or established serological profiles. "Equivocals were not included in the calculations" for the relative sensitivity/specificity and the serum characterization studies, which acts as a form of exclusion rather than an adjudication process to assign true status.
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 Enzyme-Linked Immunosorbent Assay (ELISA), which is an in vitro diagnostic test, not an AI-powered diagnostic imaging or interpretation tool that would typically involve human readers. Therefore, the concept of human readers improving with or without AI assistance is not applicable here.
6. If a standalone (i.e. algorithm only without human-in-the-loop performance) was done
Yes, the studies presented are all standalone performance evaluations of the assay itself. The results (e.g., optical density readings converted to index values) are generated by the kit components and detected instrumentally, without a human-in-the-loop interpretation step that influences the primary measurement outcome beyond standard laboratory procedures for running an ELISA.
7. The type of ground truth used
- For Relative Sensitivity and Specificity: The ground truth was based on the results of a "commercially available EIA assay" (predicate device).
- For Sensitivity and Specificity Based on Serum Characterization: The ground truth was based on serological characterization using other established EBV serologies (VCA IgG, VCA IgM) to classify samples as seronegative, acute, or seropositive. This falls under the category of established diagnostic criteria.
- For Precision, Linearity, Cross-Reactivity, Paired Sera: The ground truth was based on known characteristics of the samples used (e.g., known positive/negative samples for precision, serially diluted samples for linearity, samples with known viral antibodies for cross-reactivity).
8. The sample size for the training set
The document does not explicitly mention a "training set" in the context of machine learning or algorithm development. The device is a traditional immunoassay kit, not an AI/ML algorithm. Therefore, no specific training set data is described for this type of device. The extensive testing described in the document serves as performance validation.
9. How the ground truth for the training set was established
As there is no AI/ML algorithm and thus no explicit "training set" described, this question is not applicable. The performance characteristics were established by testing against established serological methods and clinical classifications, which serve as the reference standard or "ground truth" for validation.
§ 866.3235 Epstein-Barr virus serological reagents.
(a)
Identification. Epstein-Barr virus serological reagents are devices that consist of antigens and antisera used in serological tests to identify antibodies to Epstein-Barr virus in serum. The identification aids in the diagnosis of Epstein-Barr virus infections and provides epidemiological information on diseases caused by these viruses. Epstein-Barr viruses are thought to cause infectious mononucleosis and have been associated with Burkitt's lymphoma (a tumor of the jaw in African children and young adults) and postnasal carcinoma (cancer).(b)
Classification. Class I (general controls).