van der Groen G, Vercauteren G, Piot P
Institute of Tropical Medicine, Antwerp, Belgium.
J Virol Methods. 1987 Aug;17(1-2):35-43. doi: 10.1016/0166-0934(87)90066-8.
The enzyme-linked immunosorbent assay (ELISA) is currently being used as a sensitive screening test for HIV antibody. The immunoblot assay (IBA) and indirect immunofluorescence (IF) techniques are two recommended confirmation tests for EIA-positive sera. An indirect IF test has been developed by various laboratories using acetone fixed mixtures of uninfected and HIV-infected cells, which facilitated the reading, since nonspecific reactions were easily differentiated from specific staining. Similar results have been obtained with H9-, CEM-, and HUT78-HIV-infected and uninfected cells. Anti-nuclear antibodies and auto-antibodies resulting in false-positive EIA results, could easily be differentiated by the IF test. Aspecific fluorescence can be removed by absorption of the specimens with non-infected cells. However, IF is not suitable for the screening of large series of specimens. IF is especially well suited for quantitative analysis of serum antibody levels. Whereas serum antibody titers rise initially after infection, they decrease as AIDS develops. Heat inactivation of sera did not affect reactivity in IF, in contrast to a high rate of false-positive results obtained with heat inactivated sera in some ELISAs. A well characterized serum from an AIDS patient can be used to perform IF in order to monitor HIV infection of susceptible cells. It has been claimed that titers of neutralizing antibodies significantly correlate with the levels of IF anti-HIV antibodies. An overall correlation of 99% between IF and IBA was reported by different laboratories, when HIV ELISA-reactive European and North American sera were tested. The concordance with IBA was 97% when HIV ELISA-reactive African sera were tested.(ABSTRACT TRUNCATED AT 250 WORDS)
酶联免疫吸附测定(ELISA)目前被用作HIV抗体的敏感筛查试验。免疫印迹试验(IBA)和间接免疫荧光(IF)技术是针对ELISA阳性血清推荐的两种确证试验。不同实验室已利用未感染和HIV感染细胞的丙酮固定混合物开发出间接IF试验,这便于判读结果,因为非特异性反应很容易与特异性染色区分开来。使用H9、CEM和HUT78细胞系的HIV感染和未感染细胞也得到了类似结果。导致ELISA假阳性结果的抗核抗体和自身抗体,可通过IF试验轻松区分。非特异性荧光可通过用未感染细胞吸附标本而去除。然而,IF不适合用于大量标本的筛查。IF特别适合血清抗体水平的定量分析。感染后血清抗体滴度最初会升高,但随着艾滋病的发展会降低。与一些ELISA中热灭活血清出现的高假阳性率相反,血清的热灭活并不影响IF中的反应性。可使用一名艾滋病患者特征明确的血清进行IF试验,以监测易感细胞的HIV感染情况。据称,中和抗体滴度与IF抗HIV抗体水平显著相关。不同实验室报告称,对欧洲和北美ELISA反应性血清进行检测时,IF与IBA的总体相关性为99%。对非洲ELISA反应性血清进行检测时,与IBA的一致性为97%。(摘要截短至250词)