Detection of human immunodeficiency virus type 1 clinical isolates with reduced sensitivity to zidovudine and dideoxyinosine by RNA.RNA hybridization.
Detection of human immunodeficiency virus type 1 clinical isolates with reduced sensitivity to zidovudine and dideoxyinosine by RNA.RNA hybridization.
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通过 RNA.RNA 杂交检测对齐多夫定和双脱氧肌苷敏感性降低的人类免疫缺陷病毒 1 型临床分离株。
DOI：
10.1073/pnas.88.8.3092
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发表时间：
1991
影响因子：
11.1
通讯作者：
Crumpacker,CS
中科院分区：
文献类型：
--
作者：
Japour,AJ;Chatis,PA;Eigenrauch,HA;Crumpacker,CS
检测耐药的临床人类免疫缺陷病毒1型（HIV-1）毒株的定量快速检测仍然是一个重要的医学目标。一种结合定量RNA.RNA杂交分析的系统，测量细胞内HIV-1特异性RNA的量，已被用于检测敏感和耐药HIV-1菌株中核苷类似物的抑制水平。RNA.RNA杂交试验很容易区分以前发表的齐多夫定（ZDV; 3 '-叠氮基-3'-脱氧胸苷）耐药分离株与ZDV敏感的HIV-1分离株。ZDV对HTLV-IIIB和敏感临床HIV-1分离株的50%抑制浓度（IC 50）在0.01和0.04 μ M之间。与敏感株相比，来自3名长期ZDV治疗患者的HIV-1株显示ZDV IC 50增加了20倍以上。通过显示来自ZDV耐药分离株的HIV逆转录酶基因突变导致四个氨基酸变化（Leu-125-Trp、Ile-142-瓦尔、Thr-215-Tyr和Pro-294-Thr），包括一个变化（Thr-215-Tyr），证实了药物敏感性系统，此前已报道该变化与耐药性相关。与HTLV-IIIB相比，一种具有高水平ZDV抗性的临床HIV毒株显示2 '，3'-双脱氧肌苷IC 50增加5倍。采用RNA的药物敏感性测定。RNA杂交可用于从参加临床试验的患者中广泛筛选HIV分离株，并允许将体外耐药性与临床结果相关联。
A quantitative rapid assay to detect resistant clinical human immunodeficiency virus type 1 (HIV-1) strains remains an important medical goal. A system incorporating a quantitative RNA.RNA hybridization assay that measures the amount of intracellular HIV-1-specific RNA has been employed to detect the level of inhibition by nucleoside analogues in sensitive and resistant HIV-1 strains. The RNA.RNA hybridization assay readily distinguished previously published zidovudine (ZDV; 3'-azido-3'-deoxythymidine)-resistant isolates from ZDV-sensitive isolates of HIV-1. The 50% inhibitory concentration (IC50) of ZDV for HTLV-IIIB and sensitive clinical HIV-1 isolates is between 0.01 and 0.04 microM. HIV-1 strains from three patients on long-term ZDV therapy displayed a greater than 20-fold increase in the ZDV IC50 compared to sensitive strains. The drug sensitivity system was confirmed by showing that mutations in the HIV reverse transcriptase gene from a ZDV-resistant isolate resulted in four amino acid changes (Leu-125----Trp, Ile-142----Val, Thr-215----Tyr, and Pro-294----Thr) including one change (Thr-215----Tyr) that has been previously reported to be associated with resistance. One clinical HIV strain with high-level ZDV resistance displayed a 5-fold increase in 2',3'-dideoxyinosine IC50 compared to that of HTLV-IIIB. A drug sensitivity assay employing RNA.RNA hybridization may be useful for extensive screening of HIV isolates from patients enrolled in clinical trials and permit the correlation of in vitro resistance with clinical outcome.