乙型肝炎病毒发生rtA181T突变的临床特点与意义分析

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.     

HBV反转录酶区rtA181S变异与阿德福韦酯耐药相关性研究

目的：分析HBV反转录酶（RT）区rtA181S变异与阿德福韦酯（ADV）耐药的相关性。方法应用直接测序法筛选分析大样本慢性HBV感染者rtA181S变异的检出频率,并对1例接受ADV单药治疗失败的慢性乙型肝炎患者血清中HBV RT区基因进行克隆测序,分析相关变异形式。用XhoⅠ和SphⅠ双酶切pGEM-Teasy RT及pTriEx-HBV（C）载体后再连接,构建1.1倍HBV野生株和耐药株的重组质粒,转染人肝癌细胞系HepG2细胞,5 h后分别加入不同浓度的 ADV（0、0.033、0.100、0.330、1.000、3.300μmol/L）。隔天换药,4 d后收集细胞上清,采用实时荧光定量PCR法检测不同药物浓度作用下细胞培养上清中的HBV DNA载量,并分析其表型耐药特点。结果9830例慢性HBV感染者的12000个血清样本中,有46例样本检出rtA181S变异（单独或与其他耐药变异联合出现）,在653例中检出经典的rtN236T/A181V变异。其中随访的1例在接受ADV治疗19个月后出现了病毒学和生化学突破,直接测序检出rtA181S＋N236T变异,克隆测序分析显示在24个克隆中11个（45.83%）为野生型,6个（25.00%）为rtN236T变异型,5个（20.83%）为rtA181S变异型,1个（4.16%）为rtA181V变异型,1个（4.16%）为rtA181S＋N236T变异型。在体外实验中,rtN236T、rtA181S和rtA181S＋N236T变异株的相对复制力分别是野生株的91.35%、29.90%和68.53%。表型耐药分析显示rtN236T、rtA181S和rtA181S＋N236T变异株对ADV的灵敏性分别为野生株的1/4.41、1/3.05和1/5.43。结论 rtA181S是一种ADV耐药相关变异,可单独或联合其他变异引起患者耐药。但与经典ADV耐药变异相比,rtA181S变异引起的ADV耐药相对较弱,临床检出率较低。     

LAM耐药后ADV序贯治疗再耐药HBV变异株的动态变化

目的:观察阿德福韦酯(ADV)单药序贯治疗拉米夫定(LAM)耐药慢性乙型肝炎(CHB)出现再耐药患者HBV变异株的动态变化.方法:28例初始LAM治疗的CHB患者, 出现LAM耐药后换用ADV单药序贯治疗再次出现不充分病毒学应答或病毒学突破. 采用实时荧光定量PCR检测DNA, 分析HBV DNA动态变化; 采用直接PCR产物测序法检测耐药变异,分析变异模式变化. 对其中2例患者系列血清进行基因克隆分析其HBV耐药株动态变化.结果:28例入选患者HBV DNA载量随耐药变异株的消长而波动: 基线为(7.65±1.04)Log10copies/mL, LAM疗程中最低值中位数为3.68 Log10copies/mL; 出现病毒学突破或/和耐药变异时为(6.87±1.16) Log10copies/mL, 患者换用ADV后最低值中位数为3.78 Log10copies/mL; 患者出现耐药变异模式改变或再次病毒学突破时为(6.04±0.93) Log10copies/mL. 治疗基线及两次病毒突破3个时点HBV DNA载量均数逐渐下降, 且两两之间差异均有统计学意义. 28例患者LAM治疗后24例发生rtM204I/V伴或不伴rtL180M等变异, 4例无耐药变异; 换用ADV单药序贯治疗出现病毒学突破后, 进行测序检测耐药发现: LAM耐药变异13例(包括3例多重耐药), ADV耐药变异11例(包括3例多重耐药), 无耐药变异7例. 其中11例ADV耐药患者: rtA181V变异6例、rtA181T变异3例、rtN236T变异和rtA181V+rtN236T联合变异各1例. 对2例患者血清TA克隆发现ADV耐药准种出现和LAM耐药准种消失的过程, 同时发现对ADV耐药的rtI233M变异和对替诺福韦耐药的rtA194T变异.结论:ADV单药序贯治疗LAM耐药CHB出现再耐药患者HBV变异株动态变化存在5种形式, 且克隆分析可观察耐药株动态变化过程和发现预存的耐药准种.     

HBV rtA181T突变对Huh7细胞自噬的影响

目的探讨HBV逆转录酶区A181T(rtA181T)位点突变对Huh7细胞自噬的影响。方法 Huh7细胞分别转染1.3mer HBV野生型质粒(WT组)、1.3mer HBV rtA181T突变型质粒(181T组)及1∶1共转WT和rtA181T突变型质粒(WT+181T组)。Western Blot、免疫荧光检测细胞内HBV表面蛋白(HBs)表达水平,ELISA检测细胞外HBs Ag表达水平;Western Blot分析内源性自噬相关蛋白微管相关蛋白轻链3-Ⅱ(LC3-Ⅱ)及P62表达水平,Real-time PCR检测自噬相关基因Atg5 mRNA、Beclin1 mRNA的转录水平;同时转染绿色荧光蛋白LC3后,免疫荧光检测自噬颗粒。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果免疫荧光观察181T组、WT组及WT+181T组细胞内均有HBs表达,Western Blot结果显示181T组细胞内的HBs表达高于WT组和WT+181T组。ELISA检测3组细胞培养上清的HBs Ag水平差异有统计学意义(F=66.739,P0.001),其中181T组细胞外的HBs Ag水平(0.111±0.056)远远低于WT组(3.157±0.490)和WT+181T组(2.240±0.797)(P值均0.001);WT组、181T组和WT+181T组均可见LC3-Ⅱ自噬颗粒聚集,181T组LC3-Ⅱ表达水平高于WT组和WT+181T组,而P62水平低于WT组和WT+181T组,差异均有统计学意义(P值均0.05);181T组自噬相关基因Atg5 mRNA、Beclin1 mRNA转录水平均明显高于WT组和WT+181T组(P值均0.05)。结论 HBV rtA181T突变可导致HBs Ag分泌障碍,并增加Huh7细胞的自噬水平。     

ADV初治耐药与LAM耐药者再耐药时HBV-RT区的变异特征

目的:研究阿德福韦酯(adefovir,ADV)初治耐药和ADV用于挽救治疗拉米夫定(lamivudine,LAM)耐药者再耐药时HBV毒株RT区变异特征的差异性.方法:我院就诊的ADV初治耐药者73例,ADV用于挽救治疗LAM耐药者后再耐药者34例,共计107例,在HBVDNA突破时采用PCR方法扩增HBV-RT区基因,对PCR产物直接测序,用Chromas2.0软件分析HBV-RT区基因的核苷酸和氨基酸差异、变异模式,同时使用Clustalx1.81.msw进行基因型分析.结果:在107例患者中,存在rtA181V变异36例,rtA181T变异43例,rtN236T变异47例,rtM204I/V±rtL180M变异20例,其他相关变异如rtA181S/C、rtH/N238S/T/R/A/K、rtV214I/A、rtQ215H共12例.ADV初治组检测到rtA181V/T变异27例(36.99%)、rtN236T变异14例(19.18%)、rtA181V/T+rtN236T变异28例(38.36%),rtA181T/V和rtN236T以外的变异4例(5.48%).其中发生rtA181V和rtA181T变异患者分别有33.3%、64.5%合并rtN236T变异(P=0.0091);ADV挽救治疗LAM耐药者再耐药,检测到rtA181V/T变异21例(61.76%)、rtN236T变异2例(5.88%)、rtA181V/T+rtN236T变异3例(8.82%),rtA181T/V和rtN236T以外的变异8例(23.53%).其中发生rtA181V患者25%合并rtN236T变异,而rtA181T变异患者未检测到rtN236T变异(P=0.2311).20例(58.83%)患者合并rtM204I/V变异,这20例患者中12例(35.29%)合并rtA181V/T和/或rtN236T变异的多重耐药变异.有12例未检测到rtA181T/V和rtN236T变异,但检查到rtA181S/C(2例)、rtV214I/A(3例)、rtQ215H(1例)、rtH/N238S/T/R/A/K(6例)等ADV耐药相关变异,其中ADV挽救治疗LAM耐药患者8例,ADV初治患者4例,两组间存在统计学差异(P=0.0169).结论:ADV初治耐药主要发生rtA181V/T和rtN236T变异,ADV挽救治疗LAM耐药再耐药主要发生rtA181V/T变异,且后者耐药模式更复杂.     

HBV临床分离株复制型质粒的构建与鉴定

目的克隆HBV临床分离株全长基因组,构建真核表达复制载体,为研究HBV的耐药机制提供基础。方法从乙型肝炎患者血清中提取HBV DNA,PCR扩增HBV全长基因组,经SapI酶切后,克隆入pHY106载体,测序分析序列,然后用lipofectamine 2000将重组载体转染Huh7细胞,3 d后收集培养上清液,ELISA检测HBsAg、HBeAg水平;Real-time PCR检测细胞培养上清液中HBV含量;Southern印迹检测细胞内HBV复制中间体。结果获得5株HBV临床分离株均为野生型,其中基因B型3株,基因C型2株。成功构建5株HBV临床分离株的复制型质粒,在转染Huh7细胞的上清液中检测到HBsAg、HBeAg及HBV DNA,细胞内检测到HBV复制中间体。结论成功克隆并构建了HBV临床分离株复制型质粒,这种复制型质粒将在HBV临床分离株的耐药机制和药物敏感性等方面研究中发挥重要作用。     

乙型肝炎病毒B和C基因型全基因组的克隆与真核细胞表达

目的 构建B和C基因型重组HBV表达载体,检测其在Huh7细胞内的DNA复制和HBsAg、HBeAg的表达.方法 扩增B和C基因型HBV全基因组,并将其连接于真核表达载体pHY106,将这2个载体分别转染Huh7细胞,以pHY106空载体转染作对照.Southern印迹法检测转染72 h后HBV DNA的复制,实时定量PCR检测转染后24、48、72、96和120 h Huh7细胞内HBV DNA水平,ELISA检测转染后24、48、72、96和120 h细胞培养上清液中HBsAg和HBeAg的表达.结果 成功构建了B和C基因型HBV表达载体.转染Huh7细胞后72 h,Southern印迹法检测到细胞内HBV核心颗粒内的HBV复制中间体,包括松弛环状DNA、双链DNA和单链DNA.实时定量PCR检测发现病毒DNA复制水平可达8 lg拷贝/mL、ELISA结果显示HBsAg和HBeAg的表达于转染后72 h达高峰,然后逐渐下降.结论 成功构建B和C基因型重组HBV真核表达载体,并能在Huh7细胞内高水平复制和表达,为进一步研究HBV的结构与功能、基因表达与调控,以及抗HBV药物的筛选等提供了良好的平台.     

对阿德福韦酯自然变异耐药与抗病毒后耐药的慢性乙型肝炎患者血清HBV P基因序列比较

目的 通过对慢性乙型肝炎(CHB)患者外周血乙型肝炎病毒(HBV)聚合酶RT区序列分析,观察对阿德福韦酯(ADV)自然变异耐药及继发耐药的异同.方法 PCR产物直接测序法检测78例对ADV耐药患者外周血HBV RT区基因序列.其中曾用ADV治疗患者30例(治疗组),未抗病毒治疗48例(未治疗组),并对两组ADV相关耐药变异位点发生频数进行比较.结果 未治疗组ADV耐药的主要变异类型为rtAl81V/T 10例(20.8％),rtN/H238T/D/S 10例(20.8％)和rtV214A变异13例(27.1％);而治疗组发生的ADV耐药变异主要为rtA181V/T 16例(53.3％),没有发现rtN/H238T/D/S变异,仅发现l例rtV214A变异.结论 CHB患者中存在相当严重的ADV自然变异耐药变异,主要变异类型为rtN/H238T/D/S及rtV214A,而抗病毒治疗后该变异位点发生频数反而相对减少.rtAl81V/T变异既可以为自然变异也可以由ADV抗病毒治疗而诱发.     

Hepatitis B e antigen‐suppressing mutations enhance the replication efficiency of adefovir‐resistant hepatitis B virus strains

Summary. Hepatitis B e antigen (HBeAg) negative hepatitis B virus (HBV) infections caused by precore (PC) or basal core promoter (BCP) mutations are associated with disease progression and complications. PC or BCP mutations may enhance the replication capacity of distinct drug‐resistance‐associated polymerase mutations, but their effect on adefovir‐resistant HBV mutants is unclear. Importantly, BCP mutations were an independent risk factor for virological breakthrough in lamivudine‐resistant patients treated with adefovir. We aimed at addressing the functional consequences of PC and BCP mutations on the replication and drug susceptibility of adefovir‐resistant HBV mutants. Therefore, HBV constructs with wild type (WT) or adefovir‐resistant rtN236T, rtA181V and rtA181T mutations, with or without concomitant PC or BCP mutations, were analysed in vitro using molecular assays. The adefovir‐resistant polymerase mutations rtN236T, rtA181V and rtA181T showed a drastically reduced viral replication compared with WT. Interestingly, additional PC or BCP mutations enhanced the reduced replication efficacy of adefovir‐resistant constructs and restored HBV replication to WT level. HBV rtA181T mutants displayed abolished hepatitis B surface antigen (HBsAg) secretion, owing to a sW172* stop codon in the overlapping envelope gene. All rtN236T‐ or rtA181V/T‐containing constructs, regardless of concomitant PC or BCP mutations, were resistant to adefovir, but remained susceptible to telbivudine, entecavir and tenofovir. In conclusion, adefovir drug resistance mutations reduced viral replication, which can be significantly increased by additional HBeAg‐suppressing PC or BCP mutations. Because increased HBV replication in HBeAg‐negative patients has been associated with an unfavourable clinical course, close monitoring appears indispensable during adefovir treatment in HBeAg‐negative patients.     

The antiviral drug selected hepatitis B virus rtA181T/sW172* mutant has a dominant negative secretion defect and alters the typical profile of viral rebound

The hepatitis B virus (HBV) mutation that encodes rtA181T is selected in the viral polymerase during antiviral drug therapy and can also encode a stop codon in the overlapping surface gene at amino acid 172 (sW172*) resulting in truncation of the last 55 amino acids of the C-terminal hydrophobic region of the surface proteins. This mutation is usually detected as a mixed population with wild-type HBV. In vitro analysis revealed that the rtA181T/sW172* variant is not only defective in secretion of viral particles causing intracellular retention of surface proteins, it also has a dominant negative effect on virion but not subviral particle secretion when coexpressed with the wild type. This dominant negative effect was attributed to the truncated S protein alone. Furthermore, these truncated surface proteins were less glycosylated, and the truncated L protein was able to support virion secretion. Examination of sequential HBV DNA levels in patients failing lamivudine or adefovir therapy where only the rtA181T change was detected via polymerase chain reaction sequencing revealed that viral load rebound did not occur or was not as large as usually observed with drug-resistant HBV. CONCLUSION: The rtA181T/sW172* variant has a secretory defect and exerts a dominant negative effect on wild-type HBV virion secretion. The selection of rtA181T/sW172* reduced the typical extent of virological breakthrough, resulting in a missed diagnosis of drug resistance if viral load was used as the only criterion for drug failure, necessitating HBV polymerase chain reaction sequencing or other genotypic methods to diagnose antiviral drug resistance in these cases.     

Hepatitis B virus genotype C encoding resistance mutations that emerge during adefovir dipivoxil therapy: in vitro replication phenotype

Introduction

Hepatitis B virus (HBV) can be classified into ten genotypes (A–J), with genotypes B and C being the most common in Asia. Recent data suggest that the HBV genotype can influence disease progression, and genotype C has been associated with more aggressive liver disease than that of other genotypes. Although there is a preventative vaccine, chronic infection remains a public health problem with oral nucleos(t)ide analog therapy being the most common treatment. The HBV genome is composed of four partially overlapping reading frames, meaning that substitutions in the HBV polymerase selected during NA therapy may also alter the overlapping HBV surface antigen (HBsAg). We have recently shown that for HBV genotype D, the rtA181T/sW172stop substitution conferring resistance to adefovir dipivoxil (ADV) alters secretion of HBsAg and exerts a dominant-negative effect on wild-type virion secretion. However, the effect of this and other ADV-resistance-associated mutations on HBV replication and HBsAg secretion for the HBV genotype C, the genotype with the most severe clinical prognosis, is unknown.

Methods/Results

We constructed 1.2-mer infectious cDNA clones of HBV genotype C encoding mutations associated with ADV resistance and established an in vitro replication assay in Huh7 cells. Decreased levels of HBV DNA and HBsAg were detected for all ADV variants relative to the 1.2-mer wild-type polymerase control plasmid. Importantly, less HBsAg was detected in the cells transfected with the rtA181T resistance mutants, and the overlapping sW172stop mutation ablated secretion of HBsAg into cell culture supernatants.

Conclusions

The identification of secretion-defective HBV in the setting of ADV therapy for HBV genotype C, and to a lesser extent HBV genotype B, has major implications for the diagnosis and treatment of HBV in the Asia-Pacific region, as it is likely that quantitative HBsAg and viral load testing of serum from patients infected with HBV encoding rtA181T and rtN236T substitutions may not accurately reflect the level of replication within hepatocytes.     

逆转录酶区A181T/V和A181T/V＋N236T变异的乙型肝炎病毒感染者临床特征分析

目的：比较逆转录酶区A181T/V变异与A181T/V＋N236T变异的乙型肝炎病毒感染者临床特征的异同。方法对55例发生rtA181T/V单独变异和34例发生rtA181T/V＋rtN236T联合变异的乙型肝炎病毒感染者既往核苷（酸）类似物治疗情况进行回顾,对耐药时谷丙转氨酶、谷草转氨酶、HBV M、HBV DNA等指标进行检测,并根据耐药测序结果确定病毒的基因型。结果阿德福韦单药治疗患者发生A181T/V＋N236T变异较A181T/V变异的比例高（57.6%对42.4%,P〈0.05）,而拉米夫定换用（或加用）阿德福韦治疗患者发生 A181T/V 变异较A181T/V＋N236T变异的比例高（75.5%对24.5%,P〈0.05）;在89例患者中B基因型和C基因型分别是14例和75例,不同变异在B或C基因型中的分布无统计学差异;A181T/V变异与A181T/V＋N236T变异相比,两组患者的年龄、性别构成、谷丙转氨酶、谷草转氨酶、HBV DNA、HBsAg定量、HBeAg状态等指标均无统计学差异。结论不同的核苷（酸）类似物治疗可导致A181T/V变异和A181T/V＋N236T变异模式的差异,但这两种变异感染者临床指标无明显差异。     

Impact of hepatitis B virus rtA181V/T mutants on hepatitis B treatment failure

BACKGROUND/AIMS: Recent clinical observations reported the occurrence of amino acid substitutions at position 181 of the HBV polymerase, associated with a viral breakthrough under lamivudine or adefovir therapy. In this study, we characterized the main variants harboring the rtA181T/V mutation isolated from 10 consecutive patients who developed lamivudine and/or adefovir resistance. METHODS: We performed a clonal analysis of the HBV polymerase gene amplified by PCR from serum samples during viral breakthrough. The main mutants were then tested after transfection of Huh7 cells for their resistance profile to nucleoside analogs. RESULTS: Clonal analysis revealed the co-localization on the same HBV genome of rtA181T/V with rtN236T, but not with rtM204V/I mutations following lamivudine, adefovir or lamivudine+adefovir breakthrough. In cell culture, the rtA181T/V mutation induced a decreased susceptibility to lamivudine (<10-fold), adefovir (2- to 8-fold) and tenofovir (2- to 3-fold). Interestingly, the association of rtA181T with rtN236T on one clinical isolate genome increased the resistance to these three drugs. All the tested mutants remained sensitive to entecavir. CONCLUSIONS: Our observations suggest that a single amino acid change at position rt181 may induce cross-resistance to lamivudine and adefovir. These data emphasize the clinical relevance of genotypic and phenotypic analysis in the management of antiviral drug resistance.     

阿德福韦耐药乙型肝炎病毒株的快速检测和动态观察

目的设计PCR联合限制性片段长度多态性（PCR—RFLP）快速检测HBV阿德福韦耐药变异（rtN236T）的方法以及观察阿德福韦耐药毒株的动态变化情况。方法7例乙型肝炎患者在阿德福韦单药治疗过程中出现病毒突破或应答不完全。对其系列血清标本的HBVDNA逆转录酶部分区域进行直接或克隆后测序，设计和应用PCR—RFLP方法对rt236位点的变异情况进行检测。采用Hamming距离法计算HBV部分逆转录酶区域的基因多样性。结果l例患者出现rtA18IV变异，3例为rtN236T变异。建立了基于限制性内切酶DraI或HpaI的PCR—RFLP方法检测rtN236T变异：可以检测至少10％的弱势毒株，特异性为100％。在病毒突破前8个月可以检测到耐药毒株；该耐药毒株后来成为优势毒株。1例患者停用阿德福韦3个月后野生毒株取代耐药毒株重新成为优势株。1例患者在病毒突破后继续服用阿德福韦，rtN236T突变被一个新的突变株（rtN236V）替代。拉米夫定耐药患者的HBV逆转录酶有更明显的基因多样性。结论建立了PCR—RFLP快速检测rtN236T变异的方法；阿德福韦耐药毒株可以表现为不同的转化过程。     

Low risk of adefovir resistance in lamivudine-resistant chronic hepatitis B patients treated with adefovir plus lamivudine combination therapy: two-year follow-up

BACKGROUND/AIMS: We studied the long-term efficacy (median follow-up of 28 months) of adefovir (ADV) in combination with lamivudine (LAM) in 132 LAM-resistant Japanese patients with chronic genotype C-dominant hepatitis B virus (HBV) infection. METHODS: The viral response (undetectable HBV-DNA by PCR assay) and the predictor of viral response were evaluated. The emergence of ADV-resistant mutants was investigated during the combination therapy. RESULTS: The cumulative probability of viral response was 69% at 12 months, and 81% at 24 months. Multivariate analysis identified baseline HBe antigen status (P=0.0001), aspartate aminotransferase level (AST) (P=0.001) and HBV-DNA level (P=0.002) as determinants of viral response to treatment. At the beginning of ADV therapy, substitutions at rtA181 (rtA181T and rtA181S) were identified in 3 patients (2.3%). In the remaining 129 patients, the rtM204 mutants were identified at baseline, and two (1.6%) of the 129 patients developed new ADV-resistant mutants; one was rtA181S and another was rtA181T plus rtN236T mutation. CONCLUSIONS: Adefovir and lamivudine combination therapy effectively suppressed viral replication and maintained the efficacy well in LAM-resistant patients with chronic HBV infection. Genotypic analysis indicated that the emergence of ADV-resistant mutants is rare, at least over a period of 2 years, in patients with combination therapy.     

HBV逆转录酶区M204位点单独突变患者与M204+A181T联合突变患者临床特征的比较

目的了解HBV逆转录酶区M204位点单独突变患者与逆转录酶区M204+A181T联合突变患者临床特征的异同。方法对发生HBV逆转录酶区M204位点单独突变与逆转录酶区M204+A181T联合突变的慢性乙型肝炎患者血清HBsAg、HBVDNA和ALT水平进行统计学分析,并对所有患者的核苷(酸)类药物治疗史进行回顾性调查。结果逆转录酶区M204位点单独突变与逆转录酶区M204+A181T联合突变患者比,后者不会造成HBsAg和HBVDNA水平的下降(P值分别为0.3020和0.5917),也不会引起ALT(P=0.8001)的升高。此外,在核苷(酸)类药物应用方面,前者中单用拉米夫定的比例最高(41.38%),后者中由拉米夫定换用阿德福韦的比例最高(34.15%)。结论 rtM204位点单独突变与rtM204+rtA181T突变患者引起突变的核苷(酸)类药物使用情况有所不同,但两组病毒学指标的水平无明显的差异。