European Union Reference Laboratory for African Swine Fever (EURL), Centro de Investigación en Sanidad Animal, CISA-INIA/CSIC, Valdeolmos, Madrid, Spain.
Methods Mol Biol. 2022;2503:133-145. doi: 10.1007/978-1-0716-2333-6_9.
The present chapter describes a simple and economic indirect enzyme immunoassay (ELISA ) for African swine fever virus (ASFV) antibody detection based on the use of the soluble cytoplasmic fraction of ASFV-infected monkey stable cells (MS). The soluble antigen proteins of ASFV-infected cells are separated by sucrose precipitation centrifugation, and the supernatant above the sucrose layer is used as an ELISA antigen. The test serum sample reacts with the cytoplasmic soluble fraction, and antibodies are detected using a protein A-peroxidase conjugate. This crude antigen is currently recommended as a test reagent in screening and diagnostic tests by the World Organization for Animal Health (OIE).
本章描述了一种基于使用感染非洲猪瘟病毒(ASFV)的猴稳定细胞(MS)的可溶细胞质部分的简单且经济的间接酶联免疫吸附测定(ELISA),用于检测 ASFV 抗体。通过蔗糖沉淀离心分离感染细胞的可溶性抗原蛋白,将蔗糖层上方的上清液用作 ELISA 抗原。待检测血清样本与细胞质可溶性部分反应,然后使用蛋白 A-过氧化物酶缀合物检测抗体。目前,该粗抗原被世界动物卫生组织(OIE)推荐用于筛选和诊断检测的测试试剂。