Nielsen C M, Kvinesdal B, Vestergaard B F
Enterovirus Department, Statens Seruminstitut, Copenhagen, Denmark.
J Clin Microbiol. 1989 Jul;27(7):1609-12. doi: 10.1128/jcm.27.7.1609-1612.1989.
In the capture competition immunoassay, undiluted serum was reacted in solution with purified human immunodeficiency virus (HIV) antigen in wells of microtest plates coated with anti-HIV immunoglobulin G antibodies (HIV capture antibodies). HIV antibodies present in the serum being tested combined with the HIV antigen and thus blocked (completely or partially) the fixation of the antigen to the capture layer. Unblocked antigenic activity was measured in subsequent steps by the use of biotinylated anti-HIV immunoglobulin G and peroxidase-conjugated avidin. The assay was evaluated in comparison with indirect enzyme-linked immunosorbent assay and Western (immuno-) blot (WB). A total of 180 serum samples which reacted repeatedly as positive in indirect enzyme-linked immunosorbent assay but negative in WB were found to be negative by the capture competition assay. Of 54 serum samples showing dubious reactions (single p24 bands in WB), 53 were clearly separated into positive or negative reactions, whereas 1 serum sample gave a borderline reaction. It was concluded that a characteristic feature of this kind of inhibition assay is a very low frequency of equivocal results.
在捕获竞争免疫测定中,未稀释的血清与纯化的人类免疫缺陷病毒(HIV)抗原在包被有抗HIV免疫球蛋白G抗体(HIV捕获抗体)的微量滴定板孔中于溶液中反应。被测血清中存在的HIV抗体与HIV抗原结合,从而阻断(完全或部分)抗原与捕获层的结合。在后续步骤中,通过使用生物素化的抗HIV免疫球蛋白G和过氧化物酶偶联的抗生物素蛋白来测量未被阻断的抗原活性。该测定法与间接酶联免疫吸附测定法和蛋白质印迹法(WB)进行了比较评估。总共180份在间接酶联免疫吸附测定中反复呈阳性反应但在WB中呈阴性反应的血清样本,通过捕获竞争测定法被判定为阴性。在54份显示可疑反应(WB中出现单条p24条带)的血清样本中,53份被明确分为阳性或阴性反应,而1份血清样本给出了临界反应。得出的结论是,这种抑制测定法的一个特征是模糊结果的频率非常低。