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Diagnostic Laboratory Immunology

New Sensitive Competitive Enzyme-Linked Immunosorbent Assay Using a Monoclonal Antibody against Nonstructural Protein 1 of West Nile Virus NY99

Jiro Hirota, Yoshihiro Shimoji, Shinya Shimizu
Jiro Hirota
National Institute of Animal Health, National Agriculture and Food Research Organization, Tsukuba, Ibaraki, Japan
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Yoshihiro Shimoji
National Institute of Animal Health, National Agriculture and Food Research Organization, Tsukuba, Ibaraki, Japan
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Shinya Shimizu
National Institute of Animal Health, National Agriculture and Food Research Organization, Tsukuba, Ibaraki, Japan
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DOI: 10.1128/CVI.05382-11
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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.

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New Sensitive Competitive Enzyme-Linked Immunosorbent Assay Using a Monoclonal Antibody against Nonstructural Protein 1 of West Nile Virus NY99
Jiro Hirota, Yoshihiro Shimoji, Shinya Shimizu
Clinical and Vaccine Immunology Jan 2012, 19 (2) 277-283; DOI: 10.1128/CVI.05382-11

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New Sensitive Competitive Enzyme-Linked Immunosorbent Assay Using a Monoclonal Antibody against Nonstructural Protein 1 of West Nile Virus NY99
Jiro Hirota, Yoshihiro Shimoji, Shinya Shimizu
Clinical and Vaccine Immunology Jan 2012, 19 (2) 277-283; DOI: 10.1128/CVI.05382-11
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