Hall R A, Kay B H, Burgess G W
Immunol Cell Biol. 1987 Feb;65 ( Pt 1):103-10. doi: 10.1038/icb.1987.12.
An antigen-capture enzyme-linked immunosorbent assay (ELISA) has been developed to detect antigens of Australian flaviviruses in mosquito pools, suckling mouse brain and infected cell culture supernatant fluid. A monoclonal antibody reactive to an epitope on the envelope glycoprotein common to all flaviviruses was used as the capture antibody. Purified rabbit IgG, produced against Murray Valley encephalitis (MVE) virus, which reacted with eight Australian flaviviruses in haemagglutination inhibition (HI) and in an indirect fluorescent antibody test, was used as the indicator antibody in direct and indirect antigen-capture ELISA. A monoclonal antibody specific for a subgroup of encephalitic flaviviruses was conjugated to horseradish peroxidase and used as the indicator antibody to distinguish MVE, Kunjin and Alfuy viruses from the remainder tested. This ELISA could detect viral antigen in mosquito cell culture fluids and suckling mouse brain preparations at titres as low as 1000 TCID50/100 microliters. Viral antigen in a single mosquito infected with MVE could be detected in a pool of 500.
已开发出一种抗原捕获酶联免疫吸附测定法（ELISA），用于检测蚊群、乳鼠脑和感染细胞培养上清液中的澳大利亚黄病毒抗原。一种对所有黄病毒包膜糖蛋白上的一个表位具有反应性的单克隆抗体被用作捕获抗体。用针对墨累谷脑炎（MVE）病毒产生的纯化兔IgG作为直接和间接抗原捕获ELISA的指示抗体，该IgG在血凝抑制（HI）试验和间接荧光抗体试验中与八种澳大利亚黄病毒发生反应。一种对脑炎黄病毒亚组具有特异性的单克隆抗体与辣根过氧化物酶偶联，并用作指示抗体，以区分MVE、库京和阿尔富伊病毒与其余受试病毒。这种ELISA能够在低至1000 TCID50/100微升的滴度下检测蚊细胞培养液和乳鼠脑制剂中的病毒抗原。在500只蚊虫的混合样本中能够检测到一只感染MVE的单个蚊虫中的病毒抗原。
