title: Development and evaluation of a new epitope-blocking ELISA for universal detection of antibodies to West Nile virus.
abstract: West 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 samples 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 μl per analysis) compared 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.
authors.affiliation: Centro de Investigación en Sanidad Animal (CISA)-INIA, Ctra Algete-El Casar, s/n, 28130 Valdeolmos, Madrid, Spain.
authors.forename: Elena
authors.initials: E
authors.lastname: Sotelo
authors.forename: Francisco
authors.initials: F
authors.lastname: Llorente
authors.forename: Belen
authors.initials: B
authors.lastname: Rebollo
authors.forename: Ana
authors.initials: A
authors.lastname: Camuñas
authors.forename: Angel
authors.initials: A
authors.lastname: Venteo
authors.forename: Carmina
authors.initials: C
authors.lastname: Gallardo
authors.forename: Alison
authors.initials: A
authors.lastname: Lubisi
authors.forename: María José
authors.initials: MJ
authors.lastname: Rodríguez
authors.forename: Antonio J
authors.initials: AJ
authors.lastname: Sanz
authors.forename: Jordi
authors.initials: J
authors.lastname: Figuerola
authors.forename: Miguel Ángel
authors.initials: MÁ
authors.lastname: Jiménez-Clavero
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D019411:Clinical Laboratory Techniques / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D000939:Epitopes / Q000276:immunology; D006801:Humans; D009500:Neutralization Tests; D012680:Sensitivity and Specificity; D014714:Vertebrates; D014759:Viral Envelope Proteins / Q000276:immunology*; D014773:Virology / Q000379:methods*; D014901:West Nile Fever / Q000175:diagnosis* / Q000276:immunology / Q000662:veterinary* / Q000821:virology; D014902:West Nile virus / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D000939:Epitopes; D014759:Viral Envelope Proteins