title: All-in-one dry-reagent time-resolved immunofluorometric assay for the rapid detection of HIV-1 and -2 infections.
abstract: An all-in-one (AIO) dry-reagent time-resolved fluorometric immunoassay that requires minimal liquid handling was developed for the detection of anti-HIV-1 and -2 antibodies. To prepare the AIO wells, in vivo biotinylated capture antigens (r-Bio-HIV-1env and r-Bio-HIV-2env) were immobilized on streptavidin-coated microtitration wells and Eu(III) chelate labelled non-biotinylated tracer antigens [r-HIV-1env-Eu(III) and r-HIV-2env-Eu(III)] were dried in stable form in the same wells. The HIV AIO assay was evaluated with serum/plasma samples (n=148) from in-house and commercial panels at two different incubation times of 15 min and 1h. The overall sensitivity of the AIO assay was 98.6% and specificity was 100% for both the incubation times. The AIO assay can accept whole blood matrix. This assay is envisioned to fill the gap between the rapid point-of-care assays and traditional enzyme immunoassays (EIA) in terms of complexity and turnaround time, without compromising the performance.
authors.affiliation: Department of Biotechnology, University of Turku, Turku, Finland. Electronic address: talshe@utu.fi.
authors.forename: Sheikh M
authors.initials: SM
authors.lastname: Talha
authors.affiliation: Department of Biotechnology, University of Turku, Turku, Finland.
authors.forename: Etvi
authors.initials: E
authors.lastname: Juntunen
authors.affiliation: Department of Biotechnology, University of Turku, Turku, Finland.
authors.forename: Teppo
authors.initials: T
authors.lastname: Salminen
authors.affiliation: Faculty of Health and Medical Sciences, University of Surrey, Guildford, Surrey, United Kingdom.
authors.forename: Amninder
authors.initials: A
authors.lastname: Sangha
authors.affiliation: Department of Virology, University of Turku, Turku, Finland.
authors.forename: Tytti
authors.initials: T
authors.lastname: Vuorinen
authors.affiliation: Recombinant Gene Products Group, International Centre for Genetic Engineering & Biotechnology, Aruna Asaf Ali Marg, New Delhi, India; Translational Health Science & Technology Institute, NCR Biotech Science Cluster, Faridabad, India; Department of Paediatrics, Emory University School of Medicine, Atlanta, GA, USA.
authors.forename: Navin
authors.initials: N
authors.lastname: Khanna
authors.affiliation: Department of Biotechnology, University of Turku, Turku, Finland.
authors.forename: Kim
authors.initials: K
authors.lastname: Pettersson
mesh_terms: D001769:Blood / Q000821:virology; D015200:Fluoroimmunoassay / Q000379:methods*; D015658:HIV Infections / Q000175:diagnosis*; D015497:HIV-1 / Q000276:immunology*; D015498:HIV-2 / Q000276:immunology*; D006801:Humans; D019095:Point-of-Care Systems
keywords: All-in-one; Diagnostics; Europium; HIV; HIV antibody; Immunoassay; Testing; Time-resolved fluorometry