Sakoda Yoshihiro, Wakamoto Hiroaki, Tamura Tehpin, Nomura Takushi, Naito Michiko, Aoki Hiroshi, Morita Hiroshi, Kida Hiroshi, Fukusho Akio
Laboratory of Microbiology, Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan.
Jpn J Vet Res. 2012 Aug;60(2-3):85-94.
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.
开发了一种间接酶联免疫吸附测定(ELISA)用于筛选检测抗经典猪瘟病毒(CSFV)的抗体。通过使用抗E2蛋白的单克隆抗体进行免疫亲和纯化,从感染CSFV ALD/A76株的猪肾细胞中纯化出病毒糖蛋白,并将其吸附在微量滴定板上作为检测抗体的抗原。血清样本的每个抗体效价表示为用每个样本和阳性对照的吸光度计算出的样本与阳性值之比。该ELISA的优点是灵敏度更高:大多数中和效价超过4的血清被判定为阳性。该ELISA无法区分抗CSFV抗体和抗其他反刍动物瘟病毒的抗体,因此该ELISA中的阳性血清应通过使用CSFV、牛病毒性腹泻病毒和边界病病毒的交叉中和试验进行评估。综上所述,本研究中开发的间接ELISA是用于大规模监测经典猪瘟以检测抗CSFV抗体的有用筛选工具。