摘要
目的 乙型肝炎病毒逆转录酶（RT）区rtA181T突变与耐药相关,并可导致sW172终止突变,本研究旨在通过大样本测序与分析,明确rtA181T突变的发生特点与临床意义. 方法 研究对象为3 013例在解放军三○二医院就诊的慢性乙型肝炎患者,采集血清提取DNA,用PCR直接测序法检测耐药相关突变和病毒基因分型.构建rtA181T/sW172＊突变与野生株重组质粒分别转染HepG2细胞,比较培养上清液中HBsAg水平.HBsAg、HBV DNA水平的组间比较采用Wilcoxon秩和检验,HBV基因型构成比较采用x^2检验.结果 rtA181T的检出率为5.5％ （165/3013）,检出患者中大多数有拉米夫定和（或）阿德福韦酯用药史.突变类型上,40.0％ （66/165）为rtA181T单点突变,46.1％ （79/165）伴随阿德福韦酯耐药突变rtA181V/N236T,12.1％ （20/165）伴随拉米夫定耐药突变rtM204V/rtM204I,1.8％（3/165）伴随多重耐药突变;总体上73.9％ （122/165）在rt181位点可同时检出野生序列峰型.发生和未发生rtA181T突变的C/B基因型分别是92.1％/7.9％和82.1％/17.9％（P＜0.01）.对S蛋白影响上,98.2％ （162/165）引起sW172终止突变,1.8％（3/165）引起sW172L或sW 172S突变.体外实验中rtA181T/sW172＊突变株的HBsAg分泌受到抑制,但临床分析结果显示,检出rtA181T与未检出该突变的患者间血清HBV DNA和HBsAg水平差异无统计学意义（P＞ 0.05）. 结论 rtA181T突变与阿德福韦酯和拉米夫定用药史密切相关,可引起sW172终止突变抑制HBsAg分泌,但在患者中rt181突变株常与rt181非突变株共存,临床大样本分析结果表明单独出现该突变未对HBsAg和HBV DNA水平有明显影响.
Objective To determine the mutational profile and clinical implications of the viral reverse-transcriptase （rt）A 181T mutation in hepatitis B virus （HBV） through population-based analysis of clinical samples.Methods Serum samples from 3,013 patients who visited The 302 Hospital （Beijing,China） were investigated.HBV DNA was extracted and HBV mutations and genotypes were determined by direct sequencing.Recombinant plasmids harboring the rtA181T/sW172＊ mutant or wild type sequence were constructed and transfected into the HepG2 cell line.The levels of HBsAg in culture supernatants were compared and statistically analyzed.Results The incidence of rtA181T across the study population was 4.1％ （165/3,013）,and most of the rtAl 81T-positive patients had received adefovir and/or lamivudine.Forty percent （66/165） of the rtA 181T cases were single mutants and treatment responsive,46.1％ （76/165） included the adefovir-resistant mutation rtA 181 V/N236T,12.1％ （20/165） included the lamivudine-resistant mutation rtM204V/rtM2041,and 1.8％ （3/165） included multidrug-resistant mutations.Interestingly,73.9％ （122/165） of the rtA181T-positive samples were detected with co-existing wild-type nucleotides at the site.The rates of HBV/C to HBV/B were 92.1％ to 7.9％ in the rtA181T-positive patients,but 82.1％ to 17.9％ in the rtA181T-negative paticnts （P ＜ 0.01）.Almost all （98.2％; 129/165） of the rtA181T led to sW172＊,while only 1.8％ of the rtA181T （3/165） led to sW172L or sW172S.HBsAg secretion in vitro was reduced from the rtA181T/ sW172＊ strain,but there was no significant difference observed in the average serum HBsAg and HBV DNA levels of patients who carried or did not carry the mutant.Conclusion The HBV rtA181T mutation is closely associated with adefovir and lamivudine exposure.rtA181T may led to sW172＊,culminating in suppression of HBsAg secretion.However,co-existence of the mutant with wild-type sequences was common among our patient population,suggesting that the mutation had little impact on serum HBsAg and HBV DNA levels across the clinical study population.
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2015年第1期23-27,共5页
Chinese Journal of Hepatology
基金
国家自然科学基金（81271847、81373136）
关键词
肝炎病毒
乙型
突变
抗药性
Hepatitis B virus
Mutation
Drug resistance
