Moore J P, Wallace L A, Follett E A, McKeating J A
Department of Veterinary Pathology, University of Glasgow, UK.
AIDS. 1989 Mar;3(3):155-63. doi: 10.1097/00002030-198903000-00006.
We have developed an enzyme-linked immunosorbent assay (ELISA) specific for antibodies to the envelope glycoproteins gp120 and gp160 of HIV-1. An antibody to a conserved epitope on gp120 is adsorbed to a solid phase and used to capture gp120 and/or gp160 from solution. This may be purified recombinant protein or in simple, non-denaturing detergent extracts of different strains of HIV-1. Human serum antibodies bound to the captured antigen are subsequently detected with an anti-human antibody conjugated to alkaline phosphatase, and the AMPAK ELISA amplification system (Novo BioLabs, Cambridge, UK). With this procedure, antibodies can be detected that recognize gp120 from a wide range of divergent HIV-1 strains. The ELISA is sufficiently sensitive to detect env antibodies in sera from HIV-positive individuals at dilutions of 1:300,000. No repeatable false-positives were detected in a screen of 250 normal serum samples. Env antibodies were detected in all 37 strongly HIV-positive sera tested, and in four sera that were borderline or weakly positive in commercial ELISA. However, 55 sera positive in commercial ELISA but unconfirmable by Western blot ('ambiguously' positive) did not contain detectable env antibodies.
我们开发了一种酶联免疫吸附测定法（ELISA），专门用于检测针对HIV-1包膜糖蛋白gp120和gp160的抗体。一种针对gp120上保守表位的抗体被吸附到固相上，用于从溶液中捕获gp120和/或gp160。这可以是纯化的重组蛋白，也可以是来自不同HIV-1毒株的简单、非变性去污剂提取物。随后，用与碱性磷酸酶偶联的抗人抗体以及AMPAK ELISA扩增系统（英国剑桥的Novo BioLabs公司）检测与捕获抗原结合的人血清抗体。通过该程序，可以检测到能识别来自多种不同HIV-1毒株的gp120的抗体。该ELISA灵敏度足以检测HIV阳性个体血清中稀释度为1:300,000的env抗体。在对250份正常血清样本的筛查中未检测到可重复的假阳性。在所有检测的37份强HIV阳性血清以及4份在商业ELISA中呈临界或弱阳性的血清中均检测到env抗体。然而，55份在商业ELISA中呈阳性但无法通过蛋白质印迹法确认（“不确定”阳性）的血清中未含有可检测到的env抗体。
