title: Detection of antibodies specific to the non-structural proteins of fowl adenoviruses in infected chickens but not in vaccinated chickens.
abstract: Antibodies specific to the non-structural proteins of viruses are detected in virus-infected animals and show promise as a reliable diagnostic marker for virus infections. We examined the potential use of two non-structural proteins of fowl adenovirus (FAdV)-based, 100K and 33K, enzyme-linked immunosorbent assays (ELISAs) in the diagnosis of FAdVs. We cloned and expressed the 100K and 33K non-structural protein genes of the FAdVs in the pGEX-4T-1 plasmid vector. Purified 100K and 33K proteins alone or in combination were used as antigens in ELISAs. Antibodies specific to the 100K and 33K non-structural proteins were detected in chickens experimentally infected with FAdVs, but not in chickens vaccinated with inactivated FAdVs. In contrast, the agar gel precipitation (AGP) test detected FAdV-specific antibodies in 70.3% of the vaccinated chickens, suggesting that the non-structural protein-based ELISA could be used in the differential diagnosis of infected and vaccinated chickens. To further validate the 100K and 33K-based ELISA (100K-33K-ELISA) method, we compared its sensitivity and specificity with that of a whole virus-based ELISA and an AGP test in detecting FAdV-specific antibodies in 350 field samples. The results showed that the 100K-33K-ELISA exhibited a higher sensitivity than the AGP test and a comparable sensitivity and specificity to the whole virus ELISA. Overall, the 100K-33K-ELISA method is sensitive, specific and can be used to distinguish an acute FAdV infection from an inactivated virus-based vaccination response.
authors.affiliation: a Department of Biotechnology , Guangxi Veterinary Research Institute , Nanning , Guangxi , China.
authors.forename: Zhixun
authors.initials: Z
authors.lastname: Xie
authors.forename: Sisi
authors.initials: S
authors.lastname: Luo
authors.forename: Qing
authors.initials: Q
authors.lastname: Fan
authors.forename: Liji
authors.initials: L
authors.lastname: Xie
authors.forename: Jiabo
authors.initials: J
authors.lastname: Liu
authors.forename: Zhiqin
authors.initials: Z
authors.lastname: Xie
authors.forename: Yaoshan
authors.initials: Y
authors.lastname: Pang
authors.forename: Xianwen
authors.initials: X
authors.lastname: Deng
authors.forename: Xiuqing
authors.initials: X
authors.lastname: Wang
mesh_terms: D000256:Adenoviridae / Q000276:immunology* / Q000302:isolation & purification; D000257:Adenoviridae Infections / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary* / Q000821:virology; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D002642:Chick Embryo; D002645:Chickens / Q000276:immunology*; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D015870:Gene Expression; D005822:Genetic Vectors; D011201:Poultry Diseases / Q000175:diagnosis / Q000276:immunology* / Q000821:virology; D011993:Recombinant Fusion Proteins; D012680:Sensitivity and Specificity; D013047:Specific Pathogen-Free Organisms; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D011993:Recombinant Fusion Proteins; D017361:Viral Nonstructural Proteins