乙型肝炎基因分型同耐药位点突变的关系研究

Study on relationship between hepatitis B virus genotypes and drug resistance mutation

目的探讨耐药位点突变与乙型肝炎病毒(HBV)基因分型及其他临床信息的相关性。方法以358例慢性乙型肝炎(CHB)患者为研究对象,采用聚合酶链反应(PCR)-反向点杂交法检测耐药位点及基因分型。收集患者血清HBV DNA载量、丙氨酸氨基转移酶(ALT)水平,以及乙型肝炎病毒e抗原(HBeAg)、性别、年龄、核苷(酸)类似物用药时间等临床资料。结果所有标本均成功扩增阳性条带。HBV基因型以B型(267例)为主,其次为C型(81例)、D型(10例)。使用核苷(酸)类似物的311患者中完全野生型269例,未用药的47例患者均未见耐药突变。耐药位点突变以204及180/204位点突变为主。耐药突变与患者性别、年龄、血清HBV DNA载量、病毒基因型、ALT水平及HBeAg无明显相关性(P>0.05);用药时间越长,耐药突变率越大(P<0.05);180位点突变与204位点突变有一定相关性(P<0.05)。结论 PCR-反向点杂交技术可有效检测HBV基因型及突变位点,可有效指导临床用药。

Objective To explore correlations between hepatitis B virus （HBV）genotyping,other clinical information and drug resistance mutations.Methods 358 cases of patients with chronic hepatitis B （CHB）were selected as subjects,and the resistance loci and genotypes were detected by using the polymerase chain reaction（PCR）-reverse dot blot technique.Clinical data,such as ser-um HBV DNA loads,serum levels of alanine aminotransferase（ALT）and hepatitis B virus e antigen（HBeAg）,gender,age and length of nucleoside analogues use were collected.Results All samples were successfully amplified positive band.Type B（267 ca-ses）was the main HBV genotype,followed by type C（81 cases）and type D（10 cases）.In the 31 1 cases of patients taking nucleoside analogues,269 cases were completely wild type.No drug resistance mutation was found in 47 cases of patients not taking medicine. The drug resistance mutations mainly occured in 204 and 180/204 site.There was no significant correlation between resistance mu-tations and gender,age,serum HBV DNA loads,genotype,serum levels of ALT and HBeAg（P 〉 0.05）.While the medication time was longer,the incidence of resistant mutants was greater（P 〈0.05）.The 180 mutation had a certain correlation with 204 site mu-tation（P 〈0.05）.Conclusion PCR-reverse dot blot technology can effectively detect the HBV genotype and mutations,which could effectively guide the clinical medication.