title: Development and evaluation of indirect enzyme-linked immunosorbent assay for a screening test to detect antibodies against classical swine fever virus.
abstract: An indirect enzyme-linked immunosorbent assay (ELISA) was developed for a screening test to detect antibodies against classical swine fever virus (CSFV). Viral glycoproteins, which were purified from swine kidney cells infected with CSFV ALD/A76 strain by the immunoaffinity purification using monoclonal antibody against E2 protein, were adsorbed on a microtiter plate as the antigen for the antibody detection. Each antibody titer of serum sample was expressed as a sample per positive value calculated with optical absorbance of each sample and that of a positive control. The advantage of this ELISA is its higher sensitivity: most sera containing more than 4 neutralization titers were determined to be positive. This ELISA is unable to discriminate between antibodies against CSFV and those against other ruminant pestiviruses, therefore positive sera in this ELISA should be evaluated by a cross-neutralization test using CSFV, bovine viral diarrhea virus, and border disease virus. Taken together, the indirect ELISA developed in this study is useful screening tool to detect antibodies against CSFV for the large-scale monitoring of classical swine fever.
authors.affiliation: Laboratory of Microbiology, Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan. sakoda@vetmed.hokudai.ac.jp
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Sakoda
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Wakamoto
authors.forename: Tehpin
authors.initials: T
authors.lastname: Tamura
authors.forename: Takushi
authors.initials: T
authors.lastname: Nomura
authors.forename: Michiko
authors.initials: M
authors.lastname: Naito
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Aoki
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Morita
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kida
authors.forename: Akio
authors.initials: A
authors.lastname: Fukusho
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D001882:Border Disease / Q000097:blood / Q000175:diagnosis / Q000276:immunology; D018083:Border disease virus / Q000276:immunology; D001912:Bovine Virus Diarrhea-Mucosal Disease / Q000097:blood / Q000175:diagnosis / Q000276:immunology; D002417:Cattle; D002460:Cell Line; D006691:Classical Swine Fever / Q000097:blood / Q000175:diagnosis* / Q000276:immunology; D006692:Classical Swine Fever Virus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D001908:Diarrhea Viruses, Bovine Viral / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000662:veterinary; D009500:Neutralization Tests / Q000662:veterinary; D012680:Sensitivity and Specificity; D013552:Swine; D014759:Viral Envelope Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D014759:Viral Envelope Proteins; C087086:glycoprotein E2, classical swine fever virus