All-in-one dry-reagent time-resolved immunofluorometric assay for the rapid detection of HIV-1 and -2 infections

Sheikh M Talha; Etvi Juntunen; Teppo Salminen; Amninder Sangha; Tytti Vuorinen; Navin Khanna; Kim Pettersson

doi:10.1016/j.jviromet.2015.10.004

All-in-one dry-reagent time-resolved immunofluorometric assay for the rapid detection of HIV-1 and -2 infections

J Virol Methods. 2015 Dec 15:226:52-9. doi: 10.1016/j.jviromet.2015.10.004. Epub 2015 Oct 22.

Authors

Sheikh M Talha¹, Etvi Juntunen², Teppo Salminen², Amninder Sangha³, Tytti Vuorinen⁴, Navin Khanna⁵, Kim Pettersson²

Affiliations

¹ Department of Biotechnology, University of Turku, Turku, Finland. Electronic address: talshe@utu.fi.
² Department of Biotechnology, University of Turku, Turku, Finland.
³ Faculty of Health and Medical Sciences, University of Surrey, Guildford, Surrey, United Kingdom.
⁴ Department of Virology, University of Turku, Turku, Finland.
⁵ 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.

PMID: 26476285
DOI: 10.1016/j.jviromet.2015.10.004

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.

Keywords: All-in-one; Diagnostics; Europium; HIV; HIV antibody; Immunoassay; Testing; Time-resolved fluorometry.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Blood / virology
Fluoroimmunoassay / methods*
HIV Infections / diagnosis*
HIV-1 / immunology*
HIV-2 / immunology*
Humans
Point-of-Care Systems