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作者用胞浆斑点杂交法和逆转录酶(RT)测定法检测获得性免疫缺陷综合征患者周围血单核细胞(PBMC)培养物中的人类免疫缺陷病毒(HIV).这两种试验的主要方法如下:1.PBMC培养:用Ficoll-Hypaque密度离心法分离出HIV血清学阳性及阴性患者的PBMC,将后者用植物血凝素,(PHA)刺激3~4天,与前者混合置含20%小牛血清,2μg聚凝铵(polybrene)/ml和10%的白细胞介素2的RPMI1640培养基中共同培养数周;每周两次换液,即将培养物弃去一半,加一半新配制的培养基;7天补充一次新鲜的
The authors tested for human immunodeficiency virus (HIV) in cultures of peripheral blood mononuclear cells (PBMCs) from patients with acquired immunodeficiency syndrome by cytosolic dot blotting and reverse transcriptase (RT) assays. Both of these major Methods are as follows: 1. PBMC culture: PBMCs from HIV seropositive and negative patients were isolated by Ficoll-Hypaque densitometry. The latter were stimulated with phytohemagglutinin (PHA) for 3 to 4 days and mixed with the former 20% fetal bovine serum, 2 μg polybrene / ml and 10% interleukin 2 for several weeks; twice a week for fluid change, half of the culture is discarded and half new Preparation of medium; 7 days to add a fresh