title: Development and application of an indirect ELISA for the detection of antibodies to novel duck reovirus.
abstract: A novel duck reovirus (N-DRV) disease emerged in China in 2000 and it has become an epidemic genotype. A test for detection of virus-specific antibodies in serum samples would be useful for epidemiological investigations. Currently, Currently, serological assays for N-DRV diagnosis are not available. A test for detection of virus-specific antibodies in serum samples would be useful for epidemiological investigations. In this study, a highly sensitive and specific indirect enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to N-DRV was developed. The outer capsid (σC) of N-DRV was cloned and expressed in Escherichia coli as a coating antigen. The antigen concentration and serum dilution were optimized using a checkerboard titration. Furthermore, the specificity of σC-ELISA assay was confirmed by cross checking with other duck viral pathogens. In comparison with the western blot, the sensitivity and specificity of the σC-ELISA was 92.6% and 88.9%, respectively, and agreement of two tests was excellent with κ value of 0.786 (p < 0.05). A serological survey was performed using the assay on serum samples from different age and species of duck flocks in the Zhejiang and Jiangsu Province, China. The seropositive rate of the 1209 serum samples was 57.7%. In conclusion, the developed σC-ELISA assay is a very specific and sensitive test that will be useful for large-scale serological survey in N-DRV infection and monitoring antibodies titers against N-DRV.
authors.affiliation: Institute of Animal Husbandry and Veterinary Sciences, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Yun
authors.affiliation: College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Tai'an 271018, China.
authors.forename: Haipeng
authors.initials: H
authors.lastname: Chen
authors.affiliation: Institute of Animal Husbandry and Veterinary Sciences, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Yu
authors.affiliation: Institute of Animal Husbandry and Veterinary Sciences, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China. Electronic address: zhangcun@aliyun.com.
authors.forename: Cun
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Institute of Animal Husbandry and Veterinary Sciences, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
authors.forename: Liu
authors.initials: L
authors.lastname: Chen
authors.affiliation: Institute of Animal Husbandry and Veterinary Sciences, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Ni
authors.affiliation: Institute of Animal Husbandry and Veterinary Sciences, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
authors.forename: Jionggang
authors.initials: J
authors.lastname: Hua
authors.affiliation: Institute of Animal Husbandry and Veterinary Sciences, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
authors.forename: Weicheng
authors.initials: W
authors.lastname: Ye
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000276:immunology; D001715:Bird Diseases / Q000175:diagnosis* / Q000821:virology; D036022:Capsid Proteins / Q000276:immunology; D002681:China / Q000453:epidemiology; D004372:Ducks / Q000821:virology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D030101:Orthoreovirus, Avian / Q000276:immunology*; D012088:Reoviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D012680:Sensitivity and Specificity; D016036:Seroepidemiologic Studies; D014730:Veterinary Medicine / Q000379:methods*
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins
keywords: Indirect ELISA; Novel duck reovirus (N-DRV); Serological survey; σC protein