【摘要】 目的探索我国北京地区慢性丙型肝炎（CHC）患者直接抗病毒药物（DAA）的天然耐药突变发生情况,比较干扰素（IFN）联合利巴韦林（RBV）经治与未经治患者DAA的天然耐药突变。方法收集2009年7月^-2013年11月于北京大学第一医院就诊的101例基因1b型CHC患者,其中IFN联合RBV经治患者40例,未经治患者61例。提取患者的HCV RNA,PCR法扩增并测序HCV NS3、NS5A、NS5B区基因,进行序列比对和耐药分析,并比较天然耐药发生率。符合正态分布的计量资料组间比较采用t检验,非正态分布的计量资料组间差异比较采用Mann-Whitney U检验。计数资料组间比较采用χ²检验或Fisher精确概率法。结果共获得84例（83.17%）NS3区、92例（91.09%）NS5A区和97例（96.04%）NS5B区基因序列信息。NS3区7例（8.33%）存在DAA天然耐药突变（T54S,n=1,1.19%;R117H,n=5,5.95%;N174F,n=1,1.19%）。NS5A区19例（20.65%）存在天然耐药突变（L28M,n=7,7.61%;L31M,n=1,1.09%;P58S,n=4,4.35%;Y93H,n=7,7.61%）。NS5B区95例（97.94%）存在天然耐药突变（L159F,n=1,1.03%;C316N,n=92,94.85%;A421V,n=6,6.18%;R422K,n=1,1.03%;M426L,n=3,3.09%;V499A,n=38,39.18%）。NS3、NS5A和NS5B区天然DAA耐药突变发生率在IFN联合RBV经治与未经治患者之间差异均无统计学意义（P值均>0.05）。结论未接受过DAA治疗的我国北京地区基因1b型CHC患者中存在DAA天然耐药突变,这些突变大多为低度耐药突变,其对DAA临床疗效的影响仍需进一步研究。未观察到既往IFN联合RBV治疗对CHC患者DAA天然耐药突变率的影响。更多还原

【Abstract】 Objective To investigate the incidence of natural resistance mutations to direct- acting antiviral agents( DAAs) in patients with chronic hepatitis C( CHC) in Beijing,China,and to compare the natural resistance mutations to DAAs in interferon( IFN) / ribavirin( RBV) treatment- experienced and treatment- nave patients. Methods A total of 101 CHC patients with genotype 1b who visited Peking University First Hospital from July 2009 to November 2013 were enrolled,among whom 40 patients were once treated with IFN / RBV and 61 patients were not treated with IFN / RBV. The patients’ HCV RNA was extracted,and polymerase chain reaction was applied for amplification. The genes in HCV NS3,NS5 A,and NS5 B regions were sequenced,sequence alignment and resistance analysis were performed,and the incidence of natural resistance was compared. The t- test was applied for comparison of normally distributed continuous data between groups,and the Mann- Whitney U rank sum test was applied for comparison of abnormally distributed continuous data between groups. The chi- square test or Fisher ’s exact test was applied for comparison of categorical data between groups. Results The sequences of NS3,NS5 A,and NS5 B regions were obtained from 84( 83. 17%),92( 91. 09%),and 97( 96. 04%) patients,respectively. The natural resistance mutations to DAAs in NS3 region occurred in 7 patients( 8. 33%)( T54 S,n = 1,1. 19%; R117 H,n = 5,5. 95%; N174 F,n = 1,1. 19%),those in NS5 A region occurred in 19 patients( 20. 65%)( L28 M,n = 7,7. 61%; L31 M,n = 1,1. 09%; P58 S,n = 4,4. 35%; Y93 H,n = 7,7. 61%),and those in NS5 B region occurred in 95 patients( 97. 94%)( L159 F,n = 1,1. 03%; C316 N,n = 92,94. 85%; A421 V,n = 6,6. 18%; R422 K,n = 1,1. 03%; M426 L,n = 3,3. 09%; V499 A,n = 38,39. 18%). The incidence of resistance mutations to DAAs in NS3,NS5 A,and NS5 B regions showed no significant difference between the IFN / RBV treatment- experienced and treatment- nave patients( all P > 0. 05). Conclusion Natural resistance mutations to DAAs exist in the treatment- nave CHC patients with genotype 1b in Beijing. Most of these mutations are low resistance mutations,and their effects on the clinical efficacy of DAAs need further investigation. The association between IFN / RBV treatment and increased natural resistance mutations to DAAs was not observed in CHC patients.更多还原