Forghani B, Schmidt N J, Lennette E H
Infect Immun. 1976 Nov;14(5):1184-90. doi: 10.1128/iai.14.5.1184-1190.1976.
Hamsters immunized with human cytomegalovirus (CMV) concentrated and purified by polyethylene glycol precipitation and density gradient centrifugation produced antisera with high titers of specific viral antibody, and which showed no significant reactivity with human host cell components. The antisera had high titers of CMV antibody in complement fixation, indirect fluorescent-antibody (FA), and neutralization tests, but titers obtained by indirect radioimmunoassay (RIA) were markedly higher. The antisera were used to follow the development of CMV antigen in infected host cells by indirect RIA and indirect FA staining. Virus-specific antigen was first detectable by RIA at 8 h after infection, and by FA staining at 16 h; cells contained optimal amounts of antigen for RIA and FA assays at 72 to 100 h postinfection. Immune globulins from the antisera were labeled with 125I for use in direct RIA. The labeled globulins gave highly specific reactions with CMV-infected cells, including those infected with low-passage isolates, and showed no reactivity with cells infected with other human herpesviruses or certain other human viruses.
用聚乙二醇沉淀和密度梯度离心法浓缩纯化的人巨细胞病毒(CMV)免疫仓鼠后,产生了具有高滴度特异性病毒抗体的抗血清,且该抗血清与人宿主细胞成分无明显反应性。在补体结合试验、间接荧光抗体(FA)试验和中和试验中,抗血清具有高滴度的CMV抗体,但通过间接放射免疫测定(RIA)获得的滴度明显更高。通过间接RIA和间接FA染色,利用这些抗血清追踪感染宿主细胞中CMV抗原的发展情况。感染后8小时通过RIA首次检测到病毒特异性抗原,16小时通过FA染色检测到;感染后72至100小时,细胞含有用于RIA和FA检测的最佳抗原量。抗血清中的免疫球蛋白用125I标记,用于直接RIA。标记的球蛋白与CMV感染的细胞产生高度特异性反应,包括那些感染低传代分离株的细胞,并且与感染其他人类疱疹病毒或某些其他人类病毒的细胞无反应性。