Larder B A, Kemp S D
Molecular Sciences Department, Wellcome Research Laboratories, Beckenham, Kent, United Kingdom.
Science. 1989 Dec 1;246(4934):1155-8. doi: 10.1126/science.2479983.
Human immunodeficiency virus (HIV) isolates with reduced sensitivity to zidovudine (3'-azido-3'-deoxythymidine, AZT) from individuals with acquired immunodeficiency syndrome (AIDS) or AIDS-related complex were studied to determine the genetic basis of their resistance. Most were sequential isolates obtained at the initiation of and during therapy. Comparative nucleotide sequence analysis of the reverse transcriptase (RT) coding region from five pairs of sensitive and resistant isolates identified three predicted amino acid substitutions common to all the resistant strains (Asp67----Asn, Lys70----Arg, Thr215----Phe or Tyr) plus a fourth in three isolates (Lys219----Gln). Partially resistant isolates had combinations of these four changes. An infectious molecular clone constructed with these four mutations in RT yielded highly resistant HIV after transfection of T cells. The reproducible nature of these mutations should make it possible to develop rapid assays to predict zidovudine resistance by performing polymerase chain reaction amplification of nucleic acid from peripheral blood lymphocytes, thereby circumventing current lengthy HIV isolation and sensitivity testing.
对来自获得性免疫缺陷综合征（AIDS）或AIDS相关综合征患者的、对齐多夫定（3'-叠氮-3'-脱氧胸苷，AZT）敏感性降低的人类免疫缺陷病毒（HIV）分离株进行了研究，以确定其耐药性的遗传基础。大多数是在治疗开始时和治疗期间获得的连续分离株。对五对敏感和耐药分离株的逆转录酶（RT）编码区进行比较核苷酸序列分析，确定了所有耐药菌株共有的三个预测氨基酸取代（Asp67→Asn、Lys70→Arg、Thr215→Phe或Tyr）以及三个分离株中的第四个取代（Lys219→Gln）。部分耐药分离株具有这四种变化的组合。在RT中具有这四种突变构建的感染性分子克隆在转染T细胞后产生了高度耐药的HIV。这些突变的可重复性使得通过对外周血淋巴细胞的核酸进行聚合酶链反应扩增来开发快速检测方法以预测齐多夫定耐药性成为可能，从而规避当前冗长的HIV分离和敏感性检测。
