基因1b型慢性丙型肝炎患者DAAs天然耐药突变分析

Naturally occurring resistance mutations to direct- acting antiviral agents in chronic HCV genotype 1 b infection patients

目的探讨未经抗病毒治疗的基因1b型慢性丙型肝炎患者天然耐药突变情况。方法应用RT-PCR法扩增丙型肝炎病毒（HCV）NS3及NS5B部分片段序列，用克隆测序方法进行核苷酸序列测定。在氨基酸水平与已有报道的蛋白酶抑制剂耐药突变位点（V36、T54、V55、Q80、R155、A156、D168和V170）和聚合酶抑制剂耐药突变位点（$282、C316、S365、M414、L419、M423和Y448）进行比较。结果9例患者中3例存在直接抗病毒药物（DAAs）耐药突变，在3例检测到蛋白酶抑制剂耐药突变的患者中2例为T54S突变，1例为A156T突变，存在A156T突变的患者中同时检测到聚合酶抑制剂耐药突变M414L。结论中国DAAs初治患者中天然存在着蛋白酶抑制剂耐药突变T54S、A156T和聚合酶抑制剂耐药突蛮M414L。

Objective Investigate the naturally occurring resistance mutations to HCV protease and polymerase inhibitors in HCV genotype 1 b - infected patients who had not previously treated with direct - acting antiviral agents （DAAs）. Methods Viral sequences were analyzed in nine patients with chronic HCV genotype lb infection. Part of the NS3 and NS5B sequences were amplified by reverse transcription （RT） -PCR followed by cloned and sequenced. The sequences were compared with the resistance mutations to HCV protease and polymerasc inhibitors. The drug-resistant mutations sites examined here included eight NS3/4A protease inhibitors resistant sites （V36, T54, V55, QS0, R155, A156, D168, and V170 ）, and seven NS5B polymerase inhibitors resistant sites （$282, C316, S365, M414, L419, M423, and Y448 ）. Results Three subjects carryied the DAAs resistance mutations. These three subjects were all detected protease inhibitors resistance mutations. Of the three subjects, two had the T54S substitution,one had the A156T substitution. The subject carrying the A156T mutation were detected polymerase inhibitors resistance mutation M414L. Conclusion Protease inhibitors -resistant mutations T54S, A156T and polymerase inhibitors -resistant mutation M414L were detected in DAAs treatment -na？ ve patients in China.