Winkler Ingrid G
Methods Mol Biol. 2005;304:257-71. doi: 10.1385/1-59259-907-9:257.
Following retroviral infection, specific antibodies against viral proteins may be detected in the blood, urine, milk, or saliva of an infected animal. Enzyme-linked immunosorbent assay (ELISA) is the most commonly used method to screen for antiviral antibodies, as the technique is sensitive, reproducible, relatively inexpensive, and easily adapted to large-scale screening. This chapter describes two independent ELISA techniques. The first uses a recombinant biotinylated viral protein antigen expressed in E. coli that is purified directly on streptavidin-coated plastic wells. The second uses fixed "virus-infected" cultured cells and can be used either as a second confirmatory assay or initially in a small-scale screening to identify potentially infected individuals for further study.
在逆转录病毒感染后,可在受感染动物的血液、尿液、乳汁或唾液中检测到针对病毒蛋白的特异性抗体。酶联免疫吸附测定(ELISA)是筛查抗病毒抗体最常用的方法,因为该技术灵敏、可重复、相对便宜,且易于适用于大规模筛查。本章介绍两种独立的ELISA技术。第一种使用在大肠杆菌中表达的重组生物素化病毒蛋白抗原,该抗原直接在包被抗生物素蛋白的塑料孔上进行纯化。第二种使用固定的“病毒感染”培养细胞,可作为第二种确证测定方法,或最初用于小规模筛查,以识别潜在感染个体以便进一步研究。