title: Development and application of nsp5-ELISA for the detection of antibody to infectious bronchitis virus.
abstract: Infectious bronchitis virus (IBV) continues to be one of the most important poultry pathogens worldwide. The current commercially available enzyme-linked immunosorbent assay (ELISA) kits for IBV specific antibody detection are mostly based on the whole virion, and few serological tests based on nonstructural proteins of IBV have been developed. Herein, an alternative indirect ELISA for detection of IBV antibody was developed with IBV nonstructural protein 5 (nsp5) produced by Escherichia coli. Using an indirect immunofluorescence assay (IFA) and a commercial ELISA kit as reference, we optimized the nsp5-ELISA and determined its cut-off as 0.12. The diagnostic sensitivity (DSN), specificity (DSP) and accuracy of the nsp5-ELISA were 93.11%, 95.38% and 93.33%, respectively, compared with IFA in 660 field serum samples, and were 98.11%, 95.00% and 97.62%, respectively, compared with the commercial IBV ELISA kit (IDEXX) in 126 field sera samples. Furthermore, a time course of IBV specific antibody level detected by nsp5-ELISA following IBV infection and vaccination is consistent with that of IBV antibody detected by the commercial ELISA kit. The results presented in this study indicate that nsp5-ELISA has the potential to serve as a rapid, reliable and cost-effective method for IBV antibody detection. This study is the first to report the development of an nsp-based ELISA to detect an antibody to IBV.
authors.affiliation: Key Laboratory of Animal Virology of Ministry of Agriculture, Zhejiang University, Hangzhou 310058, PR China; Institute of Immunology and College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, PR China. Electronic address: milly0923@126.com.
authors.forename: Jing
authors.initials: J
authors.lastname: Lei
authors.affiliation: Key Laboratory of Animal Virology of Ministry of Agriculture, Zhejiang University, Hangzhou 310058, PR China. Electronic address: 2512463349@qq.com.
authors.forename: Tingting
authors.initials: T
authors.lastname: Shi
authors.affiliation: Key Laboratory of Animal Virology of Ministry of Agriculture, Zhejiang University, Hangzhou 310058, PR China. Electronic address: 446422431@qq.com.
authors.forename: Dongnan
authors.initials: D
authors.lastname: Sun
authors.affiliation: Key Laboratory of Animal Virology of Ministry of Agriculture, Zhejiang University, Hangzhou 310058, PR China. Electronic address: 1505187337@qq.com.
authors.forename: Kaikun
authors.initials: K
authors.lastname: Mo
authors.affiliation: Key Laboratory of Animal Virology of Ministry of Agriculture, Zhejiang University, Hangzhou 310058, PR China. Electronic address: yyan1512@163.com.
authors.forename: Yan
authors.initials: Y
authors.lastname: Yan
authors.affiliation: Key Laboratory of Animal Virology of Ministry of Agriculture, Zhejiang University, Hangzhou 310058, PR China. Electronic address: jinyulan@zju.edu.cn.
authors.forename: Yulan
authors.initials: Y
authors.lastname: Jin
authors.affiliation: Key Laboratory of Animal Virology of Ministry of Agriculture, Zhejiang University, Hangzhou 310058, PR China. Electronic address: liaomin4545@zju.edu.cn.
authors.forename: Min
authors.initials: M
authors.lastname: Liao
authors.affiliation: Key Laboratory of Animal Virology of Ministry of Agriculture, Zhejiang University, Hangzhou 310058, PR China; Institute of Immunology and College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, PR China. Electronic address: jyzhou@zju.edu.cn.
authors.forename: Jiyong
authors.initials: J
authors.lastname: Zhou
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000917:Antibody Formation; D002645:Chickens; D018352:Coronavirus Infections / Q000276:immunology / Q000517:prevention & control / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D001351:Infectious bronchitis virus / Q000276:immunology*; D011201:Poultry Diseases / Q000276:immunology / Q000517:prevention & control; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity; D013997:Time Factors; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins; D014765:Viral Vaccines
keywords: Enzyme-linked immunosorbent assay; Ibv; Nonstructural protein 5 (nsp5)