50例HBV基因分型及P区耐药突变分析

Analysis of HBV genotype and drug resistant mutants in HBV P gene sequences in 50 patients with hepatitis B virus

目的:调查温州乙型肝炎患者血清HBV基因分型和P区基因逆转录酶区(RT区)序列的突变情况。方法:选取50例接受核苷类似物治疗后的乙型肝炎患者作为研究对象,采用PCR产物直接测序法,并对扩增产物进行测序,将测序结果与GenBank中的标准序列进行比对分析,同时确定基因型。结果:在50例中C型44例,占88%,B型6例,占12%。存在位点突变者25例(50%)。其中检出以单位点rtM204I/V/S突变为主,9例,占36%,rtA181V/T突变4例,占16%,rtM204I/V/S+rtL180M突变5例,占20%,rtV214A,rtA181V/T+rtV173L,rtM204I/V/S+rtQ215H,rtM204I/V/S+rtL180M+rtV173L,rtM204I/V/S+rtL180M+rtV207I,rtM204I/V/S+rtL180M+rtT184A/G/I/S,rtM204I/V/S+rtL180M+rtT184A/G/I/S+rtA181V/T突变各占1例,占4%。结论:多数乙型肝炎患者在HBV P区可检出突变,突变形式多样,其中以rtM204I/V/S突变为主,应用DNA序列测定法分析HBV P区基因突变,获得的信息全面,对临床评估病情进展和实施抗病毒治疗有参考价值。

Objective：To explore the surface gene variability of hepatitis B virus（HBV） genotype and drug resistant mutants in HBV P gene sequences in Wenzhou city.Methods：The P gene from 50 HBV positive patients during nucleoside analogues therapy were amplified and sequenced directly by PCR,then these sequence results were compared with the consensus sequences in GenBank.And genotypes were determined by direct sequencing.Results：The prevalence of genotype C was 88%（44/50）,type B was 12%（6/50）.25（50%） of 50 patients in the study were mutation.Including 9 harboring rtM204I/V/S（36%）,4 harboring rtA181V/T（16%）,5 harboring rtM204I/V/S＋rtL180M（20%） and 1 harboring of rtV214A,rtA181V/T＋rtV173L,rtM204I/V/S＋rtQ215H,rtM204I/V/S＋rtL180M＋rtV173L,rtM204I/V/S＋rtL180M＋rtV207I,rtM204I/V/S＋rtL180M＋rtT184A/G/I/S,rtM204I/V/S＋rtL180M＋rtT184A/G/I/S＋rtA181V/T（4%）.Conclusion：HBV P mutations exist in most HBV patients in diverse forms.Screening HBV P gene sequences mutations by DNA,rtM204I/V/S was the most frequent mutation in patients.Sequencing could obtain rich information and thus is valuable for evaluating disease progression and administrating anti-viral therapy in clinical practice.