title: New sensitive competitive enzyme-linked immunosorbent assay using a monoclonal antibody against nonstructural protein 1 of West Nile virus NY99.
abstract: An anti-West Nile virus (anti-WNV) monoclonal antibody, SHW-7A11, was developed for competitive enzyme-linked immunosorbent assays (c-ELISAs). SHW-7A11 reacted with nonstructural protein 1 in Western blot analysis. SHW-7A11 was relatively specific for the WNV strain NY99 and recognized Kunjin and Eg101 strains in indirect ELISAs. Two c-ELISAs were developed for sera diluted 10 and 100 times and named c-ELISA10 and c-ELISA100, respectively. Both c-ELISAs detected antibodies against WNV NY99 and Kunjin strains. Little cross-reactivity was observed for antibodies against Japanese encephalitis virus and St. Louis encephalitis virus in these assays. Using the cutoff point for the St. Louis encephalitis virus, all WNV-infected chickens were found to be positive on day 21 after infection in both c-ELISAs. On the other hand, all infected chickens were found to be positive on day 35 after infection in a virus neutralization test. Our newly developed SHW-7A11-based c-ELISA can detect WNV infection with sera diluted 10 to 100 times. Therefore, this c-ELISA can be used for WNV serosurveillance of chickens and wild birds.
authors.affiliation: National Institute of Animal Health, National Agriculture and Food Research Organization, Tsukuba, Ibaraki, Japan.
authors.forename: Jiro
authors.initials: J
authors.lastname: Hirota
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Shimoji
authors.forename: Shinya
authors.initials: S
authors.lastname: Shimizu
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral; D000918:Antibody Specificity; D001715:Bird Diseases / Q000175:diagnosis* / Q000821:virology; D002645:Chickens; D003429:Cross Reactions / Q000276:immunology; D004664:Encephalitis Virus, Japanese / Q000276:immunology; D004665:Encephalitis Virus, St. Louis / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D017361:Viral Nonstructural Proteins / Q000276:immunology*; D014901:West Nile Fever / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D014902:West Nile virus / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D017361:Viral Nonstructural Proteins; C531936:nonstructural protein 1, West Nile virus