Competitive Homogeneous Immunoassay for Rapid Serodiagnosis of Hantavirus Disease.

Abstract

In this study, we describe a competitive homogeneous immunoassay that makes use of Förster resonance energy transfer (FRET) in rapid detection of pathogen-specific antibodies. The assay principle is based on competition between a monoclonal antibody (MAb) and serum antibodies to a given antigen. In the assay, named competitive FRET immunoassay (CFRET-IA), the FRET signal is induced if MAb carrying a donor label binds to an acceptor-labeled antigen. Specific antibodies in serum compete for antigen binding, resulting in reduced FRET signal. The proof-of-principle for the assay was obtained using donor-labeled Puumala virus nucleocapsid protein (PUUV-N) and acceptor-labeled anti-PUUV-N MAb. The assay was evaluated by analyzing 329 clinical samples comprising 101 from individuals with acute PUUV infection, 42 from individuals with past infection, and 186 from individuals with PUUV-seronegative sera, and the results were compared to those of reference tests. The rapid serodiagnostic test we introduced herein performed with 100% sensitivity and 99% specificity for diagnosing acute hantavirus disease.

• Authors: Satu Hepojoki, Juuso Rusanen, Jussi Hepojoki, Visa Nurmi, Antti Vaheri, Åke Lundkvist, Klaus Hedman, Olli Vapalahti
• Journal: Journal of clinical microbiology (J Clin Microbiol) Vol. 53 Issue 7 Pg. 2292-7 (Jul 2015) ISSN: 1098-660X [Electronic] United States
• PMID: 25972427 (Publication Type: Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Chemical References

• Antibodies, Viral

Topics

• Antibodies, Viral (blood)
• Fluorescence Resonance Energy Transfer
• Orthohantavirus (immunology)
• Hantavirus Infections (diagnosis)
• Humans
• Sensitivity and Specificity
• Serologic Tests (methods)
• Time Factors