Blumberg R S, Hartshorn K L, Ardman B, Kaplan J C, Paradis T, Vogt M, Hirsch M S, Schooley R T
Infectious Disease Unit, Massachusetts General Hospital, Boston 02114.
J Clin Microbiol. 1987 Oct;25(10):1989-92. doi: 10.1128/jcm.25.10.1989-1992.1987.
The detection of human immunodeficiency virus (HIV)-associated antigens was simplified by the application of dot immunobinding on a nitrocellulose matrix. Antigens were detected by applying the polyethylene glycol-precipitated supernatants of experimentally infected cultures directly onto nitrocellulose strips and sequentially incubating the strips with an anti-HIV antiserum and an alkaline phosphatase-conjugated, species-specific antiserum. The immune reaction was developed by adding the precipitable substrate indoyl phosphate. The dot immunobinding assay was nearly as sensitive as the reverse transcriptase assay in detecting HIV antigens in experimentally infected peripheral blood mononuclear cells, as well as in a T-cell line. The technique was also useful in the in vitro evaluation of antiviral agents. The dot immunobinding assay is a simple and sensitive technique that is useful in the detection of HIV antigens in studies of viral pathogenesis.
通过在硝酸纤维素膜上应用斑点免疫结合法，人类免疫缺陷病毒（HIV）相关抗原的检测得以简化。将经实验感染培养物的聚乙二醇沉淀上清液直接滴加到硝酸纤维素条上，然后依次用抗HIV抗血清和碱性磷酸酶标记的种属特异性抗血清孵育硝酸纤维素条，以此检测抗原。通过添加可沉淀底物吲哚酚磷酸来显色免疫反应。在检测经实验感染的外周血单核细胞以及T细胞系中的HIV抗原时，斑点免疫结合试验的灵敏度与逆转录酶试验相近。该技术在抗病毒药物的体外评估中也很有用。斑点免疫结合试验是一种简单且灵敏的技术，在病毒发病机制研究中检测HIV抗原时很有用。
