Das P, Hari V
Department of Biological Sciences, Wayne State University, Detroit, MI 48202.
Biotechniques. 1993 May;14(5):774-6, 778-80.
In this paper, we have used the technique known as sphere-linked immunodiagnostic assay (SLIDA) for the detection of antibodies to HIV-1 p25/24 and gp120 proteins. In this technique, the p25/24 and gp120 proteins were covalently bonded to 0.5 micron carboxylated microspheres. They were then exposed to serial dilutions of rabbit antiserum to HIV-1 p25/24, sheep antiserum to gp120 and dilutions of the corresponding normal sera. Specific immunolabeling was monitored by using gold-labeled secondary antibody. The gold particles bound to the microspheres were an indication for the presence of specific antibody in the serum sample. The gold-labeled microspheres were photographed using a transmission electron microscope, and the number of gold particles bound to the microspheres were counted manually using the electron micrographs. The specificity and sensitivity of the technique were then compared to enzyme-linked immunosorbent assay (ELISA) and Western blotting.
在本文中，我们使用了一种称为球形连接免疫诊断分析（SLIDA）的技术来检测针对HIV-1 p25/24和gp120蛋白的抗体。在该技术中，p25/24和gp120蛋白共价结合到0.5微米的羧化微球上。然后将它们分别与一系列稀释的抗HIV-1 p25/24兔抗血清、抗gp120羊抗血清以及相应的正常血清稀释液接触。通过使用金标记的二抗监测特异性免疫标记。结合到微球上的金颗粒表明血清样品中存在特异性抗体。使用透射电子显微镜对金标记的微球进行拍照，并使用电子显微照片手动计数结合到微球上的金颗粒数量。然后将该技术的特异性和灵敏度与酶联免疫吸附测定（ELISA）和蛋白质印迹法进行比较。
