中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2015年
1期
23-27
,共5页
李晓东%蒋丽红%李梵%刘妍%戴久增%赵攀%秦雅群%李进%徐东平
李曉東%蔣麗紅%李梵%劉妍%戴久增%趙攀%秦雅群%李進%徐東平
리효동%장려홍%리범%류연%대구증%조반%진아군%리진%서동평
肝炎病毒,乙型%突变%抗药性
肝炎病毒,乙型%突變%抗藥性
간염병독,을형%돌변%항약성
Hepatitis B virus%Mutation%Drug resistance
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x2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.
目的 乙型肝炎病毒逆轉錄酶(RT)區rtA181T突變與耐藥相關,併可導緻sW172終止突變,本研究旨在通過大樣本測序與分析,明確rtA181T突變的髮生特點與臨床意義. 方法 研究對象為3 013例在解放軍三○二醫院就診的慢性乙型肝炎患者,採集血清提取DNA,用PCR直接測序法檢測耐藥相關突變和病毒基因分型.構建rtA181T/sW172*突變與野生株重組質粒分彆轉染HepG2細胞,比較培養上清液中HBsAg水平.HBsAg、HBV DNA水平的組間比較採用Wilcoxon秩和檢驗,HBV基因型構成比較採用x2檢驗.結果 rtA181T的檢齣率為5.5％ (165/3013),檢齣患者中大多數有拉米伕定和(或)阿德福韋酯用藥史.突變類型上,40.0％ (66/165)為rtA181T單點突變,46.1％ (79/165)伴隨阿德福韋酯耐藥突變rtA181V/N236T,12.1％ (20/165)伴隨拉米伕定耐藥突變rtM204V/rtM204I,1.8％(3/165)伴隨多重耐藥突變;總體上73.9％ (122/165)在rt181位點可同時檢齣野生序列峰型.髮生和未髮生rtA181T突變的C/B基因型分彆是92.1％/7.9％和82.1％/17.9％(P＜0.01).對S蛋白影響上,98.2％ (162/165)引起sW172終止突變,1.8％(3/165)引起sW172L或sW 172S突變.體外實驗中rtA181T/sW172*突變株的HBsAg分泌受到抑製,但臨床分析結果顯示,檢齣rtA181T與未檢齣該突變的患者間血清HBV DNA和HBsAg水平差異無統計學意義(P＞ 0.05). 結論 rtA181T突變與阿德福韋酯和拉米伕定用藥史密切相關,可引起sW172終止突變抑製HBsAg分泌,但在患者中rt181突變株常與rt181非突變株共存,臨床大樣本分析結果錶明單獨齣現該突變未對HBsAg和HBV DNA水平有明顯影響.
목적 을형간염병독역전록매(RT)구rtA181T돌변여내약상관,병가도치sW172종지돌변,본연구지재통과대양본측서여분석,명학rtA181T돌변적발생특점여림상의의. 방법 연구대상위3 013례재해방군삼○이의원취진적만성을형간염환자,채집혈청제취DNA,용PCR직접측서법검측내약상관돌변화병독기인분형.구건rtA181T/sW172*돌변여야생주중조질립분별전염HepG2세포,비교배양상청액중HBsAg수평.HBsAg、HBV DNA수평적조간비교채용Wilcoxon질화검험,HBV기인형구성비교채용x2검험.결과 rtA181T적검출솔위5.5％ (165/3013),검출환자중대다수유랍미부정화(혹)아덕복위지용약사.돌변류형상,40.0％ (66/165)위rtA181T단점돌변,46.1％ (79/165)반수아덕복위지내약돌변rtA181V/N236T,12.1％ (20/165)반수랍미부정내약돌변rtM204V/rtM204I,1.8％(3/165)반수다중내약돌변;총체상73.9％ (122/165)재rt181위점가동시검출야생서렬봉형.발생화미발생rtA181T돌변적C/B기인형분별시92.1％/7.9％화82.1％/17.9％(P＜0.01).대S단백영향상,98.2％ (162/165)인기sW172종지돌변,1.8％(3/165)인기sW172L혹sW 172S돌변.체외실험중rtA181T/sW172*돌변주적HBsAg분비수도억제,단림상분석결과현시,검출rtA181T여미검출해돌변적환자간혈청HBV DNA화HBsAg수평차이무통계학의의(P＞ 0.05). 결론 rtA181T돌변여아덕복위지화랍미부정용약사밀절상관,가인기sW172종지돌변억제HBsAg분비,단재환자중rt181돌변주상여rt181비돌변주공존,림상대양본분석결과표명단독출현해돌변미대HBsAg화HBV DNA수평유명현영향.
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.
