title: Cloning, expression and purification of duck hepatitis B virus (DHBV) core protein and its use in the development of an indirect ELISA for serologic detection of DHBV infection.
abstract: Infecting ducks with duck hepatitis B virus (DHBV) is widely accepted as a relevant model for studying aspects of human HBV infection. However, efficient and sensitive diagnostic methods for the various infection models are limited. In order to provide a more simple and convenient method for serologic diagnosis, we improved the production of recombinant DHBV viral capsid protein (core protein) and then used it to develop an indirect enzyme-linked immunosorbent assay (ELISA) for detecting anti-DHBc antibodies (DHBcAg ELISA) in DHBV-infected ducks. Given the positive/negative cut-off value, the maximum dilution of duck sera in which anti-DHBc antibodies could be detected was 1:12,800. In addition, the DHBcAg ELISA displayed no cross reactivity with duck antisera against duck circovirus (DuCV), duck plague virus (DPV), duck hepatitis virus (DHV), duck swollen head septicemia virus (DSHSV), avian influenza virus (AIV), Riemerella anatipestifer, Salmonella anatum, or Escherichia coli. Furthermore, the coefficients of variation (CVs) of inter-assay and intra-assay experiments were both below than 10 %. When compared to PCR for accuracy on clinical samples from cases of suspected DHBV infection, the DHBcAg showed 95.45 % coincidence with PCR. In conclusion, recombinant DHBc was readily produced and used to establish a simple DHBcAg ELISA that provided a highly specific and sensitive method for analysis of clinical samples.
authors.affiliation: Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, People's Republic of China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Liu
authors.forename: Renyong
authors.initials: R
authors.lastname: Jia
authors.forename: Mingshu
authors.initials: M
authors.lastname: Wang
authors.forename: Juan
authors.initials: J
authors.lastname: Huang
authors.forename: Dekang
authors.initials: D
authors.lastname: Zhu
authors.forename: Shun
authors.initials: S
authors.lastname: Chen
authors.forename: Zhongqiong
authors.initials: Z
authors.lastname: Yin
authors.forename: Yin
authors.initials: Y
authors.lastname: Wang
authors.forename: Xiaoyue
authors.initials: X
authors.lastname: Chen
authors.forename: Anchun
authors.initials: A
authors.lastname: Cheng
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral / Q000097:blood; D003001:Cloning, Molecular; D004372:Ducks; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D018347:Hepadnaviridae Infections / Q000175:diagnosis / Q000662:veterinary / Q000821:virology; D015611:Hepatitis B Virus, Duck / Q000235:genetics / Q000378:metabolism*; D006524:Hepatitis, Viral, Animal / Q000175:diagnosis / Q000821:virology; D016133:Polymerase Chain Reaction; D011201:Poultry Diseases / Q000097:blood / Q000175:diagnosis / Q000821:virology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014758:Viral Core Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D014758:Viral Core Proteins