Liu J, Yue J, Wu S, Yan Y
Department of Clinical Laboratory, Shanghai Pulmonary Hospital, Shanghai, China.
Arch Virol. 2007;152(10):1799-805. doi: 10.1007/s00705-007-1019-9. Epub 2007 Jul 6.
The database of genotypic drug resistance mutations in HIV-1 subtype B circulating in developed industrial countries has been well established; however, little is known regarding the prevalence of genotypic resistance patterns in patients harboring non-subtype-B HIV-1 variants in most Asian countries.
To characterize the polymorphisms and emergence of drug-resistance mutations, resistance to antiretroviral drugs in naïve and pretreated patients infected with HIV-1 CRF01_AE isolates in Fujian province, China.
HIV-1 pol amplicons from 52 pre- and 14 post-treatment samples were obtained by reverse transcription-polymerase chain reaction (RT-PCR) and sequencing. All of the 14 antiretroviral-treated patients were under a fixed regimen of stavudine (d4T), lamivudine (3TC) and nevirapine (NVP), and they had been on treatment for a mean of 6 months (SD, 4 months). The sequence data were analyzed using the Bioedit software, and the data regarding drug resistance mutations were obtained using the Stanford software ( http://hivdb.stanford.edu ).
In comparison with the consensus sequence of B strains, the most common protease polymorphisms in HIV-1 CRF01_AE strains prevailing in Fujian Province, China, were I13V (76.9%), E35D (76.9%), M36I (100%), R41K (98.1%), H69K (90.4%), and L89M (96.2%). Protease mutations between CRF01_AE strains and B' variants prevailing in China were observed. The proportion of substitutions L63P, A71T/V, V77I and I93L in subtype B' sequences was considerably higher than in CRF01_AE viruses, while the proportion of L10I, M36I and K20R/I substitutions in subtype B' sequences was relatively lower than in CRF01_AE strains. A high level of resistance to nucleoside reverse transcriptase inhibitors (NRTIs) (28.6%, 4/14) and non-nucleoside reverse transcriptase inhibitors (NNRTIs) (35.7%, 5/14) was found in treatment-experienced patients. High-level resistance to nevirapine (NVP) and lamivudine (3TC) was found in the stavudine/lamivudine/nevirapine (d4T/3TC/NVP) treatment regimen. The overall drug resistance rate was 42.9% (6/14), the resistance rates to two and to all three drugs under treatment were 14.3% (2/14) and 7.1% (1/14), respectively.
This study is the first report on polymorphisms and emergence of drug-resistance mutations in HIV-1 subtype CRF01_AE prevailing in China. These findings provide useful information on global HIV genetic variability and non-B drug resistance.
在发达国家流行的HIV-1 B亚型中，基因型耐药突变数据库已经建立完善；然而，在大多数亚洲国家，携带非B亚型HIV-1变异株的患者中基因型耐药模式的流行情况却知之甚少。
对中国福建省感染HIV-1 CRF01_AE毒株的初治和经治患者中抗逆转录病毒药物耐药性的多态性和耐药突变的出现情况进行特征分析。
通过逆转录聚合酶链反应（RT-PCR）和测序，从52份治疗前和14份治疗后样本中获取HIV-1 pol扩增子。14例接受抗逆转录病毒治疗的患者均采用司他夫定（d4T）、拉米夫定（3TC）和奈韦拉平（NVP）的固定治疗方案，平均治疗时间为6个月（标准差4个月）。使用Bioedit软件分析序列数据，并使用斯坦福软件（http://hivdb.stanford.edu）获取耐药突变数据。
与B毒株的共有序列相比，在中国福建省流行的HIV-1 CRF01_AE毒株中，最常见的蛋白酶多态性为I13V（76.9%）、E35D（76.9%）、M36I（100%）、R41K（98.1%）、H69K（90.4%）和L89M（96.2%）。观察到CRF01_AE毒株与在中国流行的B'变异株之间的蛋白酶突变情况。B'亚型序列中L63P、A71T/V、V77I和I93L替换的比例明显高于CRF01_AE病毒，而B'亚型序列中L10I、M36I和K20R/I替换的比例相对低于CRF01_AE毒株。在有治疗经验的患者中发现对核苷类逆转录酶抑制剂（NRTIs）（28.6%，4/14）和非核苷类逆转录酶抑制剂（NNRTIs）（35.7%，5/14）的高水平耐药。在司他夫定/拉米夫定/奈韦拉平（d4T/3TC/NVP）治疗方案中发现对奈韦拉平（NVP）和拉米夫定（3TC）的高水平耐药。总体耐药率为42.9%（6/14），对治疗中两种药物和三种药物的耐药率分别为14.3%（2/14）和7.1%（1/14）。
本研究是关于中国流行的HIV-1 CRF01_AE亚型多态性和耐药突变出现情况的首次报告。这些发现为全球HIV基因变异性和非B亚型耐药性提供了有用信息。
