Shafer R W, Levee D J, Winters M A, Richmond K L, Huang D, Merigan T C
Division of Infectious Diseases, Stanford University Medical Center, California, USA.
J Clin Microbiol. 1997 Feb;35(2):520-2. doi: 10.1128/jcm.35.2.520-522.1997.
Human immunodeficiency virus type 1 (HIV-1) pol mutations are responsible for HIV-1 resistance to current antiretroviral drugs. HIV-1 RNA extraction with QIAamp HCV kit spin columns (Qiagen, Chatsworth, Calif.) followed by reverse transcription-PCR successfully recovered a 1,008-bp pol fragment from the plasma of 31 of 34 HIV-1-infected patients that was suitable for sequencing and recombinant-virus studies. The minimum HIV-1 RNA concentration required for gene recovery was 30 to 40 copies/ml, which was similar to the minimal HIV-1 RNA concentration required when phenol-chloroform or silica beads are used for RNA extraction.
1型人类免疫缺陷病毒(HIV-1)的pol基因突变是导致HIV-1对当前抗逆转录病毒药物产生耐药性的原因。使用QIAamp HCV试剂盒旋转柱(Qiagen公司,加利福尼亚州查茨沃思)提取HIV-1 RNA,随后进行逆转录-聚合酶链反应,成功地从34例HIV-1感染患者中的31例患者血浆中回收了一段1008 bp的pol片段,该片段适用于测序和重组病毒研究。基因回收所需的最低HIV-1 RNA浓度为30至40拷贝/毫升,这与使用苯酚-氯仿或硅胶珠进行RNA提取时所需的最低HIV-1 RNA浓度相似。