安徽淮北市CHB患者HBV基因型、耐药类别分布与耐药突变位点的分析

Analysis of HBV genotypes,drug resistant distribution and mutation sites in CHB patients in Huaibei

目的了解安徽淮北市慢性乙型肝炎(CHB)患者乙型肝炎病毒(HBV)基因型、耐药类别分布特征及耐药位点。方法选取2018年1月—2020年12月淮北矿工总医院及其下属医院收治的HBV-DNA阳性CHB患者141例,收集血清。采用荧光定量PCR反应进行HBV-DNA定量检测。采用PCR-RFLP法检测HBV基因分型。采用Sanger法测序分析P区基因序列核苷类似物耐药相关突变位点和耐药情况。结果141例CHB患者中有109例成功分型,其中HBV基因型以C型为主(71.56%),B型(17.43%)次之,B+C型最少(11.01%)。CHB患者耐药突率为25.69%,且C型(28.21%)最高。常用核苷类似物的耐药性,以拉米夫定及阿德福韦最高(60.71%),替比夫定次之(25.00%)、恩替卡韦最低(3.57%)。P区检出16种耐药突变类型和3种突变模式,均以C型为主。其中阿德福韦以rtN236T和rtA181T/S常见;拉米夫定以rtL180M+M204I/V为主,rtA181T及V207M次之;恩替卡韦以rtA181S+T184N为主;替比夫定以rtL180M+M204I/V常见。结论安徽淮北市HBV基因型以C型为主,C型耐药突变率最高。核苷类似物耐药均以C型为主,且突变模式较复杂。不同类型CHB患者耐药突变类别及位点存在差异性,因此有必要对HBV基因型及耐药相关突变进行检测,以提高核苷类似物抗病毒治疗的疗效。

Objective To analyze the genotypes,drug resistant distribution and drug mutation sites of hepatitis B virus(HBV)in chronic hepatitis B(CHB)patients in Huaibei,Anhui province.Methods A total of 141 CHB patients with HBV-DNA positive who were admitted to Huaibei General Miners Hospital and their branch hospitals from January 2018 to December 2020 were selected and their serum samples were collected.Their HBV-DNA was quantified by fluorescence quantitative PCR.Their HBV genotypes were determined by PCR-RFLP method.The drug-resistant sites in the P region of HBV gene were analyzed by Sanger sequencing.Results Among the 141 CHB patients,109 patients were successfully classified,where genotype C accounted for the most(71.56%),followed by genotype B(17.43%),and genotype B+C was the lowest(11.01%).The drug-resistance mutation rate of these CHB patients was 25.69%,where type C(28.21%)was the highest.The drug resistance rate of nucleoside analogs was highest in lamivudine and adefovir(60.71%),followed by telbivudine(25.00%),and entecavir was the lowest(3.57%).About 16 drug resistance mutation types and 3 mutation patterns were detected in the P region,most of which were type C.Among them,adefovir was mainly rtN236T and rtA181T/S;lamivudine was mainly rtL180M+M204I/V,followed by rtA181T and V207M;entecavir was mainly rtA181S+T184N;telbivudine was mostly rtL180M+M204I/V.Conclusions The genotype of CHB patients in Huaibei is mainly genotype C.The drug-resistant mutation rate of genotype C is higher than that of genotypes B and B+C.The resistant type of nucleoside analogs are mainly genotype C,and the mutation pattern is relatively complex.Patients with different HBV genotypes have different drug-resistance mutation sites,so it is necessary to detect HBV genotypes and related drug-resistance mutations in CHB patients during treatment with nucleoside analogs,so as to improve the efficacy of antiviral therapy.