(270 days)
For the qualitative and semi-quantitative determination of IgG antibodies to Epstein-Barr Virus (recombinant) Early Antigen Diffuse (EBV-EA-D IgG) in human serum by indirect enzyme immunoassay. The Is-EBV-EA-D IgG Test Kit may be used in combination with other Epstein-Barr serologies (Viral Capsid Antigen (VCA) IgG and IgM , Epstein-Barr Nuclear Antigen-1 (EBNA-1) IgG and IgM, Early Antigen-Diffuse (EA-D) IgM and heterophile antibody as an aid in the diagnosis of infectious mononucleosis (IM). The evaluation of paired sera, to determine a significant increase in EA-D IgG antibody titer, can also aid in the diagnosis of acute infection. These reagents can be used either manually or in conjunction with the MAGO® Plus Automated Processor.
The Is-EBV-EA-D IgG ELISA Kit is an enzyme-linked immunosorbent assay (ELISA) for the detection of IgG to Epstein Barr Early Antigen diffuse in human serum. Recombinant EA-D antigen is bound to microwells. Diluted patient sera, Cut-Off Calibrator and controls are placed in the microwells and incubated. Anti-EA-D IgG antibodies, if present, will bind to the antigen forming antigen-antibody complexes. Residual sample is eliminated by aspirating, Conjugate (horseradish peroxidase-labeled anti-human IgG) is added and will bind to these complexes. Unbound conjugate is removed by aspiration and washing. Substrate is then added and incubated. In the presence of bound enzyme the substrate is converted to an end product. The absorbance of this end product can be read spectrophotometrically at 450 nm (reference 600-630 nm) and is directly proportional to the concentration of IgG antibodies to EA-D present in the sample.
The provided document describes the "Is-EBV-EA-D IgG ELISA Kit" and its performance characteristics, primarily for establishing substantial equivalence to a predicate device.
Here's an analysis of the acceptance criteria and study findings based on the provided text:
1. Table of Acceptance Criteria and Reported Device Performance
The document does not explicitly state pre-defined acceptance criteria for the "Is-EBV-EA-D IgG ELISA Kit" to meet. Instead, it presents performance characteristics and concludes substantial equivalence based on a comparison with a predicate device. The values reported for the proposed device are presented as its performance, without explicit thresholds set beforehand as "acceptance criteria."
However, we can infer performance metrics that were evaluated:
| Performance Characteristic | Reported Device Performance (Is-EBV-EA-D IgG ELISA Kit) |
|---|---|
| Relative Sensitivity (Late Infection) | 81.8% |
| Relative Sensitivity (Current Infection) | 28.1% |
| Relative Specificity (Convalescent) | 89.7% |
| Relative Specificity (Seronegative) | 100.0% |
| Overall Agreement | 79.9% |
| Intra-assay Precision (Manual) | 2.07-13.88% CV (Positive samples, all sites collectively, range across samples) |
| Intra-assay Precision (MAGO Plus) | 1.77-18.00% CV (Positive samples, all sites collectively, range across samples) |
| Inter-assay Precision (Manual) | 5.12-9.08% CV (Positive samples, all sites collectively, range across samples) |
| Inter-assay Precision (MAGO Plus) | 5.09-11.61% CV (Positive samples, all sites collectively, range across samples) |
| Cross-reactivity | No cross-reactivity expected with Varicella Zoster, Cytomegalovirus, and Herpes Simplex Virus. (Tested with 16 sera, 15 anti-VZV IgG positive, 3 anti-CMV IgG positive, 3 anti-HSV positive, all non-reactive for anti-EA-D IgG) |
| Correlation (Manual vs. MAGO Plus) | Pearson Correlation Coefficient: 0.989 (Based on 193 serum samples) |
2. Sample Size Used for the Test Set and Data Provenance
- Sample Size for Test Set: 184 frozen retrospective sera for clinical sensitivity and specificity.
- Data Provenance: The sera were retrospectively collected and "characterized as research frozen retrospective sera." The document does not specify the country of origin, only stating it was tested by "an independent clinical commercial laboratory." It is retrospective data.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of Those Experts
The document does not specify that "experts" were used to establish the ground truth in the traditional sense of a human review panel. Instead, the sera were "characterized" based on a combination of different EBV serologies.
- No explicit number of experts is mentioned.
- No qualifications of experts are provided.
4. Adjudication Method for the Test Set
There was no adjudication method described. The "ground truth" (EBV serological status) was established by characterizing the sera based on results from other EBV serologies (VCA IgG, VCA IgM, EBNA IgG, heterophile antibody), not through a human consensus or adjudication process for the test device itself.
5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study
No MRMC comparative effectiveness study was done. This device is an in-vitro diagnostic (IVD) assay (ELISA), which measures biochemical markers and thus does not involve human readers interpreting images or data alongside an AI.
6. Standalone (Algorithm Only) Performance
Yes, the study primarily describes the standalone performance of the "Is-EBV-EA-D IgG ELISA Kit" as an in-vitro diagnostic test. There is no "human-in-the-loop" component described for interpreting the ELISA results themselves beyond the technician performing the test and reading the absorbance values to calculate an index value, which then falls into predefined interpretative ranges (negative, equivocal, positive). The "MAGO Plus Automated Processor" is mentioned as an automated way to run the assay, but it automates the physical assay steps, not interpretive steps.
7. Type of Ground Truth Used
The ground truth used was EBV Serological Status, established by testing the sera with a panel of other established EBV serologies (VCA IgG, VCA IgM, EBNA IgG, heterophile antibody). This is a form of reference standard data based on established diagnostic tests for Epstein-Barr Virus infection. It is not pathology, expert consensus on the device's output, or outcomes data.
8. Sample Size for the Training Set
The document does not describe a "training set" in the context of an algorithm or machine learning device. This is an ELISA kit, a traditional biochemical assay. Therefore, there is no explicit training set as would be found in AI/ML device development. The assay's parameters would have been developed and optimized internally by the manufacturer, but this is not typically referred to as a "training set" in this context.
9. How the Ground Truth for the Training Set Was Established
As there is no described training set for an algorithm, the concept of establishing ground truth for a training set does not apply directly. The development of the ELISA kit itself relies on established serological principles and reagents (e.g., recombinant EA-D antigen), with internal validation studies conducted during development to optimize its performance, but this is distinct from establishing ground truth for an AI/ML training set.
§ 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).