title: Development of blocking ELISA for detection of antibodies against H9N2 avian influenza viruses.
abstract: A blocking enzyme-linked immunosorbent assay (bELISA) was developed for detection of antibodies against H9N2 avian influenza viruses (AIVs) based on a monoclonal antibody specific to the hemagglutinin (HA) protein of H9N2 AIV. The specificity of the bELISA was tested using antisera against H3, H4, H5, H7, and H10 AIVs and other avian viruses. The average percent inhibition (PI) value of 116 non-immune serum samples from ducks and specific pathogen-free (SPF) chickens was 2.80%. The selected cut-off PI values for negative and positive sera were 17.6% and 25.0%, respectively. A high correlation of >96% was identified between the bELISA and hemagglutinin inhibition (HI) assay, according to the detection results of sera from infected chickens (n=30) and from chickens vaccinated with an inactivated H9N2 vaccine (n=40). Sera collected from vaccinated chickens and ducks (n=660) at different weeks post vaccination that were positive by the HI assay were confirmed with the bELISA. The results revealed a time-dependent increase in antibody levels. Therefore, the bELISA offers the potential advantage of a high throughput, rapid, sensitive, and specific method for detection of specific antibodies against H9N2.
authors.affiliation: Innovation Team for Pathogen Ecology Research on Animal Influenza Virus, and Department of Avian Infectious Disease, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China.
authors.forename: Jianmei
authors.initials: J
authors.lastname: Yang
authors.affiliation: Innovation Team for Pathogen Ecology Research on Animal Influenza Virus, and Department of Avian Infectious Disease, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China.
authors.forename: Xiaoguang
authors.initials: X
authors.lastname: Dai
authors.affiliation: Innovation Team for Pathogen Ecology Research on Animal Influenza Virus, and Department of Avian Infectious Disease, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China.
authors.forename: Hongjun
authors.initials: H
authors.lastname: Chen
authors.affiliation: Innovation Team for Pathogen Ecology Research on Animal Influenza Virus, and Department of Avian Infectious Disease, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China.
authors.forename: Qiaoyang
authors.initials: Q
authors.lastname: Teng
authors.affiliation: Innovation Team for Pathogen Ecology Research on Animal Influenza Virus, and Department of Avian Infectious Disease, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China.
authors.forename: Xuesong
authors.initials: X
authors.lastname: Li
authors.affiliation: Innovation Team for Pathogen Ecology Research on Animal Influenza Virus, and Department of Avian Infectious Disease, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China.
authors.forename: Guangyu
authors.initials: G
authors.lastname: Rong
authors.affiliation: Innovation Team for Pathogen Ecology Research on Animal Influenza Virus, and Department of Avian Infectious Disease, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China.
authors.forename: Liping
authors.initials: L
authors.lastname: Yan
authors.affiliation: Innovation Team for Pathogen Ecology Research on Animal Influenza Virus, and Department of Avian Infectious Disease, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China.
authors.forename: Qinfang
authors.initials: Q
authors.lastname: Liu
authors.affiliation: Innovation Team for Pathogen Ecology Research on Animal Influenza Virus, and Department of Avian Infectious Disease, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China. Electronic address: lizejun@shvri.ac.
authors.forename: Zejun
authors.initials: Z
authors.lastname: Li
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D002645:Chickens; D004372:Ducks; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006385:Hemagglutination Inhibition Tests; D053127:Influenza A Virus, H9N2 Subtype / Q000276:immunology*; D007252:Influenza Vaccines / Q000008:administration & dosage / Q000276:immunology*; D005585:Influenza in Birds / Q000175:diagnosis* / Q000821:virology; D012680:Sensitivity and Specificity; D015164:Vaccines, Inactivated / Q000008:administration & dosage / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D007252:Influenza Vaccines; D015164:Vaccines, Inactivated
keywords: Antibodies; Avian influenza virus; Blocking ELISA; H9N2 subtype