核苷和核苷酸类药物经治和未治慢性乙型肝炎患者中HBV rtV191I/sW182*突变分析

Analysis of rtV191I/sW182* mutation in chronic hepatitis B patients treated with nucleos/tide analogues

目的研究耐药相关rtV191I突变导致的sW182*型S蛋白截短突变在核苷和核苷酸类药物(NAs)经治和未治慢性乙型肝炎(CHB)患者中检出情况。方法从本课题组数据库选择592条来自于HBVDNA和HBsAg均阳性CHB患者的HBV逆转录酶和相应S蛋白序列,NAs经治和未治患者分别为318例和274例。用生物信息学方法分析HBV基因型和rtV191I/sW182*突变,雅培I 2000全自动化学发光免疫分析仪及相应试剂检测含突变标本血清学指标。结果 rtV191I/sW182*突变检出率在NAs经治组和未治组分别为6.29%(20/318)和1.46%(4/274),两组差异有统计学意义(P=0.003)。此突变可在5种治疗方案组检出,分别为拉米夫定(LAM)换用阿德福韦酯(ADV)组(3/26,11.54%)、ADV单药组(8/93,8.60%)、替比夫定单药组(1/15,6.67%)、LAM单药组(7/110,6.36%)和LAM单药治疗后加用ADV组(1/17,5.88%),但在恩替卡韦相关的7种治疗方案组中均未检出(0/28,0%)。rtV191I/sW182*突变检出组患者的年龄和NAs经治比例均显著高于未检出组[年龄:(47.96±13.33)岁vs(37.86±13.74)岁,P=0.001;NAs经治比例:83.33%vs 52.46%,P=0.003]。该突变仅在C基因型患者中检出(24/474,5.06%)。含rtV191I/sW182*突变血清标本以HBeAg阳性为主,可检测到滴度不同的HBsAg(18.6～106552IU/ml)。结论 rtV191I/sW182*突变与NAs经治和年龄显著相关,LAM和ADV治疗组检出率较高,检出者年龄较高;仅在C基因型中检出。突变株生物学和致病特点有待进一步研究。

Abstract： Objective To detect nucleos/tide analogues （NAs） resistance-induced hepatitis B virus （HBV） S protein truncated mutation sW182＊ related to rtV191I mutation in NAs-treated and untreated chronic hepatitis B （CHB） patients. Methods From our laboratory database, 592 sequences of the HBV reverse transcriptase and the related S protein were obtained from CHB patients （318 of NAs-treated and 274 untreated） positive in both serum HBV DNA and HBsAg. Bioinformatics analysis was done to determine the HBV genotype and the rtV191I/sW182＊ mutation. HBV serological markers with rtV191I/sW182＊ mutation were assayed by chemi- luminescent microparticle immunoassay （CMIA） using ARCHITECT I 2000 analyzer （Abbott Diagnostics, North Chicago, IL）. Results The detection rates of rtV191I/sW182＊ mutation in NAs-treated group （20/ 318, 6.29%） is much higher that in the untreated group （4/274, 1.46% ; P=0. 003）. This mutation couldbe detected with five NAs treatment regimes, L e. , lamivudine （LAM） and adefovir dipivoxil （ADV） se- quential, therapy （3/26, 11.54%）, ADV monotherapy （8/93, 8.60%）, telbivudine monotherapy （1/15, 6.67%）, LAM monotherapy （7/110, 6.36%） and LAM to LAM and ADV add-on therapy （1/17, 5.88%）, except seven various entecavir-related therapies （0/28, 0% ）. The average age and the ratio of NAs treated patients in the rtV191I/sW182＊ mutation group were significantly higher than that in the wild typegroup （age： 47.96±13.33 vs37.86±13.74, P=0.001; ratio： 83.33% vs 52.46%, P=0.003） .Allmu- tant HBV strains were categorized into genotype C （24/474, 5.06%） , Samples with mutant HBV showed different serum HBsAg levels （18.6 106 552 IU/ml） and most of them were HBeAg positive. Conclusions The carboxyl terminal truncation of the HBV surface （S） protein might increase its pathogenicity. The rtV191I/sW182＊ mutation is related to the average age and the NAs treatment of the patients. The mutation was detected only in CHB patients with genotype C HBV infection. The patients with this mutation were elder in age and LAM and ADV might increase the possibility of this mutation.