摘要
目的了解辽宁省自2004年至2008年治疗前人类免疫缺陷病毒1型（HIV-1）感染者中耐药基因突变新变化及HIV—1耐药株的流行率。方法应用抽签法选取经确诊但未接受过治疗的HIV-1感染者血浆样本20份，提取RNA。通过病毒载量检测，HIV-1蛋白酶（PR）全长和部分反转录酶（RT）基因区通过逆转录PCR（RT．PCR）法和巢式PCR法得到扩增样本。经测序，运用ContigExpress对序列拼接、编辑和校正，上传耐药序列至斯坦福大学HIV耐药数据库（http：／／hivdb．stanford．edu）进行分析，寻找耐药位点，评价目前未治疗感染者中病毒基因耐药新变化和耐药株流行趋势。结果20例未治疗确诊感染者血浆中，13份病毒载量检测值大于1000拷贝／ml的样本得到有效扩增，经测序得到65条有效序列。耐药基因型分析发现2个在辽宁省新出现的亚型H和CRF10-CD。13份样本均未发现核苷类反转录酶抑制剂（NRTIs）相关突变，4份存在非核苷类反转录酶抑制剂（NNRTIs）相关耐药突变，突变位点分别为P225H、K238S、V179D、K238T，突变率为30．8％（4／13）；1份（7．7％）样本蛋白酶抑制剂（PI）主要相关耐药位点154S发生突变，发生PI多重交叉耐药。3份样本仅在PR区分别有1处次要耐药突变，突变位点分别为LIOV、A71V，但对蛋白酶基因耐药没有任何影响。结论辽宁省HIV-1耐药基因型发现新亚型，PR区基因发现主要耐药突变，应考虑启用二线蛋白酶抑制剂药物。
Objective 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. Methods RNA was extracted from 20 plasma samples of diagnosed untreated HIV-l-infected treatment-naive patients by drawing method. After the viral loading （VL） test, the protease and nucleoslde 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）. Results 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） CRFIO 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 154S 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. Conclusion 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.
出处
《中华预防医学杂志》
CAS
CSCD
北大核心
2009年第11期951-955,共5页
Chinese Journal of Preventive Medicine
