Competitive Homogeneous Immunoassay for Rapid Serodiagnosis of Hantavirus Disease
2015 (English)In: Journal of Clinical Microbiology, ISSN 0095-1137, E-ISSN 1098-660X, Vol. 53, no 7, 2292-2297 p.Article in journal (Refereed) Published
In this study, we describe a competitive homogeneous immunoassay that makes use of Forster 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.
Place, publisher, year, edition, pages
2015. Vol. 53, no 7, 2292-2297 p.
Microbiology in the medical area
IdentifiersURN: urn:nbn:se:uu:diva-260637DOI: 10.1128/JCM.00663-15ISI: 000358287700039PubMedID: 25972427OAI: oai:DiVA.org:uu-260637DiVA: diva2:847804
This study was supported by the following: the Finnish Funding Agency for Technology and Innovation (TEKES) (grant 440118), the Academy of Finland (grants 1122539 and 1259376), the Sigrid Juselius Foundation (grant 4703777), the Foundation for Finnish Inventions, an Integrative Life Science doctoral program, the Otto A. Malm Foundation, and the Foundation for Clinical Chemistry Research.2015-08-212015-08-212015-08-21Bibliographically approved