Davis G R, Blumeyer K, DiMichele L J, Whitfield K M, Chappelle H, Riggs N, Ghosh S S, Kao P M, Fahy E, Kwoh D Y
Salk Institute Biotechnology/Industrial Associates, San Diego, California.
J Infect Dis. 1990 Jul;162(1):13-20. doi: 10.1093/infdis/162.1.13.
Eighty-six peripheral blood mononuclear cell (PBMC) samples from 30 patients with AIDS were analyzed using a transcription-based amplification system (TAS) and the polymerase chain reaction (PCR). Human immunodeficiency virus type 1 (HIV-1) sequences were detected by amplification-mediated hybridization in 98% of the samples, 52% of which were positive for p24 antigen by ELISA. Neither TAS (93%) nor PCR (95%) detected HIV-1 sequences in all 86 samples. The hybridization-detection methods (slot blot, bead-based sandwich, and solution) used to detect the HIV-1-specific TAS products had a clear influence on the efficiency of detecting and quantitating the levels of HIV-1 present in these samples. The reproducibility of amplification of constant amounts of HIV-1 RNA and beta-globin DNA by TAS and PCR was studied over 3 months. The results indicated that variations of 10- and 5-fold in the HIV-1 sequence levels could be detected between samples by TAS and PCR, respectively. Within the range of sensitivities for each assay used, the administration of zidovudine did not appear to reduce the amount of HIV-1 nucleic acid sequences as observed in PBMC obtained serially from six AIDS patients.
使用基于转录的扩增系统（TAS）和聚合酶链反应（PCR）对30例艾滋病患者的86份外周血单个核细胞（PBMC）样本进行了分析。通过扩增介导的杂交在98%的样本中检测到了1型人类免疫缺陷病毒（HIV-1）序列，其中52%的样本通过ELISA检测p24抗原呈阳性。TAS（93%）和PCR（95%）均未在所有86份样本中检测到HIV-1序列。用于检测HIV-1特异性TAS产物的杂交检测方法（狭缝印迹、基于磁珠的夹心杂交和溶液杂交）对检测和定量这些样本中HIV-1水平的效率有明显影响。在3个月的时间里研究了TAS和PCR对恒定数量的HIV-1 RNA和β-珠蛋白DNA扩增的可重复性。结果表明，TAS和PCR分别可以在样本之间检测到HIV-1序列水平10倍和5倍的变化。在每种检测方法的灵敏度范围内，对从6例艾滋病患者连续获取的PBMC进行观察，齐多夫定的使用似乎并未减少HIV-1核酸序列的量。
