title: Development and validation of an indirect enzyme-linked immunosorbent assay for the detection of antibodies against duck swollen head hemorrhagic disease virus.
abstract: An indirect enzyme-linked immunosorbent assay (iELISA) was developed for detecting antibody to duck swollen head hemorrhagic disease virus (DSHDV) using purified whole virus by sucrose density gradient ultracentrifugation as a coating antigen. Antiserum against DSHDV strains HY-99 (hyperimmume serum) was prepared in 30-day-old ducks by vaccination with inactivated DSHDV and used as positive sera. The iELISA test was optimized with different reagents or dilutions. The validation results showed that this assay was only specific for antibodies against duck viral swollen head hemorrhagic disease. The OD450 value for positive serum diluted 1:800 was also determined to be greater than the positive threshold. The highest coefficient of variation value was 3.66% for the intra-assay and 5.79% for the interassay, which were all less than 10%. This assay has been successfully applied to the examination of the duck sera clinically. These results in this study indicate that the newly-developed iELISA offers a precise, specific, sensitive, and reproducible means of measuring DSHDV antibodies in duck sera.
authors.affiliation: Avian Disease Research Center, College of Veterinary Medicine of Sichuan Agricultural University, Yaan, Sichuan 625014, China.
authors.forename: Chuanfeng
authors.initials: C
authors.lastname: Li
authors.forename: Anchun
authors.initials: A
authors.lastname: Cheng
authors.forename: Mingshu
authors.initials: M
authors.lastname: Wang
authors.forename: Na
authors.initials: N
authors.lastname: Zhang
authors.forename: Chanjuan
authors.initials: C
authors.lastname: Shen
authors.forename: Jinlong
authors.initials: J
authors.lastname: Yang
authors.forename: Dekang
authors.initials: D
authors.lastname: Zhu
authors.forename: Renyong
authors.initials: R
authors.lastname: Jia
authors.forename: Qihui
authors.initials: Q
authors.lastname: Luo
authors.forename: Xiaoyue
authors.initials: X
authors.lastname: Chen
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D004372:Ducks; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D011201:Poultry Diseases / Q000097:blood / Q000175:diagnosis* / Q000276:immunology / Q000517:prevention & control; D012087:Reoviridae / Q000276:immunology*; D012088:Reoviridae Infections / Q000097:blood / Q000276:immunology / Q000517:prevention & control / Q000662:veterinary*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014765:Viral Vaccines / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D014765:Viral Vaccines