title: Performance characteristics of the immunoglobulin G-capture BED-enzyme immunoassay, an assay to detect recent human immunodeficiency virus type 1 seroconversion.
abstract: Recently, we developed an immunoglobulin G (IgG)-capture BED-enzyme immunoassay (BED-CEIA) to identify recent human immunodeficiency virus (HIV) type 1 (HIV-1) seroconversion for use in incidence estimates. We have established an algorithm for its use; developed quality control reagents to monitor the assay; and evaluated its performance for interrun, intrarun, and operator variability. Analysis of 144 individual plates, which involved multiple plate lots and several operators over more than a year, indicated that the coefficients of variation (CVs) were between 10 and 15% for raw optical density (OD) values in the dynamic range between 0.5 and 2.0 OD units; the CVs decreased to 5 to 10% when the OD was normalized (OD-n; OD-n = specimen OD/calibrator OD). The intrarun CVs were generally in the range of 5 to 10% for specimens with ODs <0.5 and less than 5% for specimens with ODs >0.5. The level of concordance between multiple plate lots (n = 6) and multiple operators (n = 7) was quite high (R(2) > 0.9). Comparison of the results of the initial and the confirmatory tests with specimens with OD-n values </=1.5 demonstrated a high degree of correlation (R(2) = 0.92); 566 (92%) of 615 of specimens tested in the two modes retained the same classification (recent or long-term infection). The values for those specimens with changed classifications (n = 49) were close to the cutoff (OD-n = 1.0), as expected. The twofold difference in the HIV IgG contents between the controls and the calibrator reagents was exploited to monitor individual plate runs by using a control plot, which was incorporated into the spreadsheet for data entry and run monitoring. This information provides baseline data for the successful transfer of BED-CEIA to other laboratories and the use of BED-CEIA for the detection of recent HIV seroconversion and the calculation of incidence estimates worldwide.
authors.affiliation: HIV Immunology and Diagnostics Branch, Mailstop D12, Centers for Disease Control and Prevention, 1600 Clifton Rd., Atlanta, GA 30333, USA.
authors.forename: Trudy
authors.initials: T
authors.lastname: Dobbs
authors.forename: Susan
authors.initials: S
authors.lastname: Kennedy
authors.forename: Chou-Pong
authors.initials: CP
authors.lastname: Pau
authors.forename: J Steven
authors.initials: JS
authors.lastname: McDougal
authors.forename: Bharat S
authors.initials: BS
authors.lastname: Parekh
mesh_terms: D015483:HIV Antibodies / Q000097:blood*; D006679:HIV Seropositivity; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000097:blood*
chemical_list: D015483:HIV Antibodies; D007074:Immunoglobulin G