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Development and evaluation of a new epitope-blocking ELISA for universal detection of antibodies to West Nile virus

AuthorsSotelo, Elena; Llorente, Francisco; Rebollo, Belén; Camuñas, Ana; Venteo, Ángel; Gallardo, Carmina; Lubisi, Alison; Rodríguez, María José; Sanz, Antonio J.; Figuerola, Jordi ; Jiménez-Clavero, Miguel A.
KeywordsWest Nile virus
Antibody detection
Epitope-blocking ELISA
Monoclonal antibody
Wild birds
Issue DateJun-2011
CitationJournal of Virological Methods 174 (2011) 35–41
AbstractWest Nile virus (WNV) is an emerging zoonotic pathogen with a wide range of hosts, including birds, horses and humans. The development and evaluation of the performance of a new enzyme-linked immunosorbent assay (ELISA) are described for rapid detection of WNV-specific antibodies in sam- ples originating from an extensive range of vertebrates susceptible to WNV infection. The assay uses a monoclonal antibody (MAb) which binds whole virus particles and neutralizes infection in vitro by recognizing a neutralizing epitope within the envelope (E) glycoprotein of the virus. This MAb, labelled with horseradish peroxidase, was used to compete with WNV-specific serum antibodies for virus-binding in vitro. The epitope-blocking ELISA was optimized in a manner that enabled its validation with a number of experimental and field sera, from a wide range of wild bird species, and susceptible mammals. The new ELISA exhibited high specificity (79.5–96.5%) and sensitivity (100%), using the virus-neutralization test as reference standard. It also required a much lower volume of sample (10 pl per analysis) com- pared to other ELISAs available commercially. This new method may be helpful for diagnosis and disease surveillance, particularly when testing samples from small birds, which are available in limited amounts
Publisher version (URL)http.//dx.doi.org/10.1016/j.jviromet.2011.03.015
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