Wu Shao-hui, Lu Chun-ming, Jiang Feng-xia, E Shuang, Ma Ning, Liang Xue-wei, Gai Xiao-qun
Department of Acquired Immunodeficiency Sydrome and Sexually Transmitted Disease, Liaoning Centers for Disease Control and Prevention, Shenyang 110005, China.
Zhonghua Yu Fang Yi Xue Za Zhi. 2009 Nov;43(11):951-5.
To investigate the HIV-1 drug resistance associated mutations and examine the susceptibility of HIV-1 with these mutations to antiretroviral in treatment-naive individuals in Liaoning province from 2004 to 2008.
RNA was extracted from 20 plasma samples of diagnosed untreated HIV-1-infected treatment-naive patients by drawing method. After the viral loading (VL) test, the protease and nucleoside reverse transcriptase coding regions were amplified by RT-PCR, nested PCR and sequence analysis directly. Levels of resistance and prevalence were evaluated according to the Stanford University HIV Drug Resistance Database's algorithm (http://hivdb.stanford.edu).
Among the 20 plasma samples, 13 got PCR products because of their VL values higher than 1000 copies/ml.Meanwhile, the 13 samples got 65 sequences by using 5 primers each. Polymorphisms in subtype H and circulating recombinant forms (CRFs) CRF10_CD sequences were identified. An overall prevalence of 30.8% (4/13) resistance to NNRTIs, 7.7% (1/13) to PI and no NRTIs mutations were found. The most frequent substitutions (4/13) in the RT region at positions P225H, K238S, V179D, K238T and a major position I54S in PR implied to a multiple drug-resistance. A71V or L10V only, respectively, substitution in PR was found in 3 samples, but no any worse with drug sensitivity.
HIV-1 polymorphisms in subtype H and CRFs CRF10_CD sequences were identified circulating in Liaoning. A major mutation position I54S in PR implied that it would be the time to commence a higher level drug regimen.
调查2004年至2008年辽宁省初治HIV-1感染者中与HIV-1耐药相关的突变,并检测携带这些突变的HIV-1对抗逆转录病毒药物的敏感性。
采用抽取法从20例确诊未治疗的HIV-1感染初治患者的血浆样本中提取RNA。进行病毒载量(VL)检测后,通过RT-PCR、巢式PCR和直接序列分析扩增蛋白酶和核苷类逆转录酶编码区。根据斯坦福大学HIV耐药数据库的算法(http://hivdb.stanford.edu)评估耐药水平和流行率。
20份血浆样本中,13份因VL值高于1000拷贝/ml获得PCR产物。同时,这13份样本每份使用5对引物共获得65条序列。鉴定出H亚型和循环重组型(CRF)CRF10_CD序列中的多态性。对非核苷类逆转录酶抑制剂(NNRTIs)的总体耐药率为30.8%(4/13),对蛋白酶抑制剂(PI)的耐药率为7.7%(1/13),未发现核苷类逆转录酶抑制剂(NRTIs)突变。RT区最常见的替换(4/13)发生在P225H、K238S、V179D、K238T位点,PR区的一个主要位点I54S提示多重耐药。3份样本中PR区分别仅发现A71V或L10V替换,但药物敏感性无明显下降。
在辽宁发现H亚型和CRF CRF10_CD序列中的HIV-1多态性在流行。PR区的一个主要突变位点I54S提示应开始采用更高水平的药物治疗方案。