黑龙江省未经治疗的艾滋病患者蛋白酶和反转录酶基因型耐药性检测与分析

Study of genotypic resistance mutations of protease and reverse transcriptase in non-therapeutic acquired immune deficiency syndrome patients in Heilongjiang Province

目的 对黑龙江省流行的HIV-1毒株的蛋白酶基因及反转录酶基因进行基因型耐药分析,为开展大规模临床抗病毒治疗提供本底数据.方法 采用套式-PCR法扩增黑龙江省内49例HIV-1感染者及AIDS患者外周血单个核细胞中前病毒cDNA的部分蛋白酶和反转录酶基因,分析基因序列,与国际耐药数据库比对,分析耐药变异.结果 49例患者HIV毒株亚型分析为B＇亚型.在扩增出蛋白酶基因的47份标本中,未发现对蛋白酶抑制剂的原发突变,但发现大量对蛋白酶抑制剂的次要突变,V771占91.5%、L63P占76.6%、I93L占74.5%、E35D占61.7%、R57K占19.1%、R41K占10.6%、A71V占8.5%、M36I占8.5%、L10I占6.4%、D60E占6.4%、L89M占4.2%和G16E占2.1%.在扩增出反转录酶基因的44份标本中,1份有反转录酶抑制剂的原发突变M184I,占2.3%,均有反转录酶抑制剂的次要突变,I135L/T/R/V占81.8%、V106I占22.7%、V179D/E占11.4%、R211K占9.1%、L214F占4.5%、V189I占4.5%和V108I占2.3%.结论 目前在黑龙江省未接受抗反转录病毒治疗的患者中未发现对蛋白酶抑制剂耐药的原发突变,耐药突变处于低水平,但应及时进行耐药突变检测,防止多重耐药和交叉耐药的出现和流行.

Objective To study the presence of drug resistant mutations of protease and reverse transcriptase among human immunodeficiency virus （HIV）-1 strains isolated from treatment naive HIV/ acquired immune deficiency syndrome （AIDS） patients in Heilongjiang Province of China and to provide the baseline data for starting antiretroviral therapy in this area. Methods The protease and reverse transcriptase gene sequences were amplified by nested-polymerase chain reaction （PCR） and then sequenced. The results were compared to the subtype B consensus amino acid sequence and analyzed with Stanford HIV-db drug resistance sequence interpretation. Results The results showed that HIV strains from 49 patients were classified as subtype B＇. No primary mutations associated with protease inhibitor were detected. Some secondary mutations associated with protease inhibitor were detected, which included V77I（91.5%）, L63P（76.6%）, I93L（74.5%）, E35D（61.7%）, R57K （19.1%）, R41K（10.6%）, A71V（8.5%）, M36I（8.5%）, L10I（6.4%）, D60E（6.4%）, L89M （4.2%） and G16E（2. 1%）. Only one case had a primary mutation M184I that was associated with resistance to reverse transcriptase inhibitors. However, many secondary mutations associated with resistance to reverse transcriptase inhibitors were found, including I135L/T/R/V（81.8%）, V106I（22.7%）, V179D/E（11.4%）, R211K（9.1%）, L214F（4.5%）, V189I（4.5%） and V108I（2. 3%）.Conclusions The prevalence of genotypic anti-HIV drug resistance is very low in treatment naive HIV/AIDS patients in Heilongjiang Province. However, closely monitoring on drug resistance mutation is very important for preventing the development and prevalence of multi-drug resistant or cross drug resistant HIV.