HIV/HCV合并感染者和HCV单纯感染者NS3/4A蛋白酶、NS5A抑制剂的天然耐药变异分析

Naturally occurring resistance-associated variants to NS3/4A protease and NS5A inhibitors in patients with HIV/HCV co-infection or HCV infection alone

目的通过对抗病毒治疗前HIV/HCV合并感染者和HCV单纯感染者HCV NS3/4A蛋白酶、NS5A抑制剂相关耐药位点进行检测,探讨上述患者天然耐药变异的差异。方法收集2016年1月—2020年1月在广州医科大学附属市八医院住院或门诊就诊的246例HIV/HCV合并感染者和HCV单纯感染者的血清标本,使用Illumina二代测序平台进行测序,比较已在中国获批准的NS3/4A蛋白酶、NS5A抑制剂相关耐药变异在两组患者的差异,纳入分析的药物包括阿舒瑞韦/达拉他韦(ASV/DCV,基因1b型),艾尔巴韦/格拉瑞韦(EBR/GZR,基因1b型)和格卡瑞韦/哌仑他韦(GLE/PIB,泛基因型)。符合正态分布的计量资料两组间比较采用t检验,不符合正态分布的计量资料两组间比较采用Mann-Whitney U检验。计数资料两组间比较采用χ2检验或Fisher精确检验。结果本研究纳入的246例患者的血清样本中,239例(97.2%)成功进行PCR扩增并测序,包括102例HIV/HCV合并感染者和137例HCV单纯感染者。对ASV/DCV和EBR/GZR相关耐药变异分析结果提示:HCV 1b型的HIV/HCV合并感染者和HCV单纯感染者中存在Y56F、Q80K/L、S122N/R/T等ASV和GZR相关耐药变异,以及L31M、Y93H等DCV和EBR相关耐药变异;2例HIV/HCV合并感染者(均为Y56F+Y93H)和2例HCV单纯感染者(Q80L+L31M和Y56F+Y93H)对EBR/GZR双靶点同时存在耐药变异,两组患者各耐药变异差异均无统计学意义(P值均>0.05)。对泛基因型的GLE/PIB相关耐药变异分析结果提示,仅在HCV 3a型的患者中,存在3例PIB相关耐药变异,分别为HIV/HCV合并感染者中2例Y93H耐药变异和HCV单纯感染者中1例Y93H耐药变异(P=0.590),所有纳入分析的患者均不存在GLE/PIB双靶点耐药变异。结论HIV/HCV合并感染者和HCV单纯感染者的NS3/4A蛋白酶、NS5A抑制剂相关天然耐药变异无明显差异。

Objective To investigate the difference in naturally occurring resistance-associated variants(RAVs)between the patients with HIV/HCV co-infection and those with HCV infection alone by detecting the drug resistance loci associated with HCV NS3/4A protease and NS5A inhibitors.Methods A total of 246 patients with HIV/HCV co-infection or HCV infection alone who were hospitalized or attended the outpatient service in Guangzhou Eighth People’s Hospital,Guangzhou Medical University,from January 2016 to January 2020 were enrolled in this study.Serum samples were collected and next-generation sequencing(Illumina platform,PE250)was used for sequencing.The two groups of patients were compared in terms of RAVs associated with NS3/4A protease and NS5A inhibitors approved in China,and the drugs for analysis included asunaprevir/daclatasvir(ASV/DCV)and elbasvir/grazoprevir(EBR/GZR)for HCV genotype 1b and glecaprevir/pibrentasvir(GLE/PIB)for pan-genotypes.The t-test was used for comparison of normally distributed continuous data between two groups,and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups;the chi-square test or the Fisher’s exact test was used for comparison of categorical data between two groups.Results Among the 246 serum samples included in this study,239 samples(97.2%)were successfully amplified by PCR and sequenced,with 102 samples from the patients with HIV/HCV co-infection and 137 from the patients with HCV infection alone.The analysis of RAVs associated with ASV/DCV and EBR/GZR showed that Y56F,Q80K/L,and S122N/R/T associated with ASV and GZR and L31M and Y93H associated with DCV and EBR were observed in patients with HIV/HCV(genotype 1b)co-infection or HCV(genotype 1b)infection alone;2 patients with HIV/HCV co-infection had the RAVs of Y56F+Y93H associated with EBR/GZR,and 2 with HCV infection alone had the RAVs of Q80L+L31M and Y56F+Y93H,respectively,associated with EBR/GZR,with no significant difference in RAVs between the two groups(both P>0.05).The analysis of RAVs associated with GLE/PIB for pan-genotypes showed that 3 patients with PIB-associated Y93H RAV were observed among the patients with HCV genotype 3a infection,among whom 2 had HIV/HCV co-infection and 1 had HCV infection alone(P=0.590),and in addition,no RAVs associated with GLE/PIB were observed.Conclusion There is no significant difference in naturally occurring RAVs associated with HCV NS3/4A protease and NS5A inhibitors between the patients with HIV/HCV co-infection and those with HCV infection alone.