Andreoni M, Sarmati L, Parisi S G, Ercoli L, Rocchi G
Department of Public Health and Cellular Biology, University of Rome Tor Vergata, Italy.
J Med Virol. 1992 Nov;38(3):207-13. doi: 10.1002/jmv.1890380310.
A semimicromethod was established for isolating human immunodeficiency virus (HIV) in plasma using 48-well plates and a pool of peripheral blood mononuclear cells (PBMC) from several donors as targets for infection, which increases the efficiency of isolation by reducing the effect of variability due to diverse donor cell susceptibility to HIV infection. The addition of H9 cells to the PBMC cultures did not affect measurable titers. Nevertheless, it potentiated strongly virus replication in terms of p24 production in the supernatant of the wells with HIV isolates, thus facilitating interpretation of the results. The titration of a virus strain of a known titre and reverse transcriptase activity in parallel provided a constant parameter of efficiency and reproducibility within each experiment, permitting comparison with results from other laboratories. The reproducibility of the method was highly significant (r = 0.97, P < 0.001); 68% of the 22 plasma samples from HIV-infected individuals tested by this method were positive. The presence of plasma HIV titer correlated well (P < 0.02) with the low count of CD4+ cells of less than 300/mm3, but not with the presence of the p24 antigen in the serum.
建立了一种半微量方法，用于在血浆中分离人类免疫缺陷病毒（HIV），该方法使用48孔板和来自多个供体的外周血单个核细胞（PBMC）池作为感染靶标，通过减少由于不同供体细胞对HIV感染的易感性差异所产生的变异性影响，提高了分离效率。在PBMC培养物中添加H9细胞不影响可测量的滴度。然而，就含有HIV分离株的孔的上清液中p24的产生而言，它强烈增强了病毒复制，从而便于结果的解释。同时对已知滴度和逆转录酶活性的病毒株进行滴定，为每个实验提供了效率和可重复性的恒定参数，便于与其他实验室的结果进行比较。该方法的可重复性非常显著（r = 0.97，P < 0.001）；用该方法检测的22份来自HIV感染者的血浆样本中，68%呈阳性。血浆HIV滴度的存在与CD4 +细胞计数低于300/mm3密切相关（P < 0.02），但与血清中p24抗原的存在无关。
