乙型肝炎病毒感染不同状态下血清病毒反转录酶区准种特点及其临床意义  

The characterization and clinical significance of quasispecies in the reverse transcriptase region of hepatitisB virus in hepatitis B virus infected patients with different disease stages

作　　者：姚碧莲[1] 刘峰[2] 黄素园[2] 于德敏[2] 陈立[2] 韩悦[2] 李新华[2] 张欣欣[2] 

机构地区：[1]上海交通大学医学院附属瑞金医院特需医疗保健中心200025 [2]上海交通大学医学院附属瑞金医院感染科,200025

出　　处：《中华传染病杂志》2011年第12期717-722,共6页Chinese Journal of Infectious Diseases

基　　金：国家“十二五”重大专项（2012ZXl0002007002-003）;中国肝炎防治基金会光辉基金项目（GHF2010210）;国家自然科学基金资助项目（30872251）

摘　　要：目的研究HBV感染不同状态下血清病毒反转录酶（RT）区准种特点和临床意义。方法50例未接受抗病毒治疗的HBV感染者分为慢性HBV携带者（ASC）组10例、慢性乙型肝炎（CHB）组30例和乙型肝炎肝硬化（LC）组lO例。收集患者外周血血清，抽提HBVDNA，PCR扩增RT区基因组后克隆、测序。每例患者获得15～30个序列，进行HBVRT区的准种异质性分析及相关基因突变统计等生物信息学分析。多均数比较采用方差分析，中位数比较采用非参数检验分析，非计量资料比较采用y。检验。结果总共测序1221个克隆，其中ASC组152个，CHB组780个，LC组289个。3组问基因型构成差异无统计学意义。准种复杂度为LC组〉CHB组〉ASC组，3组间差异有统计学意义（F--33，400，P〈0，05）。准种离散度为LC组〉CHB组〉ASC组，I，C组与CHB组及ASC组均差异有统计学意义（F--18．070，P％0．05），CHB组与ASC组间差异无统计学意义。结论慢性HBV感染过程中，免疫清除期比免疫耐受期具有更宽的HBV变异谱系，随着病程的延长和病情加重，HBV的准种趋向复杂。Objective To characterize the profile and clinical significance of hepatitis B virus （HBV） quasispecies in patients infected with hepatitis B virus based on the sequence of reverse transcriptase （RT） region. Methods Fifty HBV infected treatment naive patients were enrolled and divided into three groups, asymptomatic carriers （ASC） group （10 cases）, chronic hepatitis B （CHB） group （30 cases） and liver cirrhosis （LC） group （10 cases）. HBV genomes were extracted from serum samples. The sequence of RT region was amplified by polymerase chain reaction （PCR） and cloned into vectors. Fifteen to thirty clones per sample were selected, sequenced and analyzed by bioinformatics software. The mean values among groups were compared by analysis of variance. The median values among groups were compared by nonparametric statistics. The enumeration data were analyzed by X2 test. Results Totally 1221 HBV RT region nucleotide sequences were obtained （152 from ASC patients, 780 from CHB patients and 289 from LC patients）. Genotype distribution showed no difference among three groups. However, the quasispecies complexity showed significantdifferences among the three groups, LC group 〉CHB group〉ASC group （F= 33. 400, P〈0.05）. The quasispecies diversity was LC group 〉CHB group〉 ASC group, and that of LC group was significantly different from the other two groups （F= 18. 070, P〈0.05）, while there was no significant difference between CHB and ASC patients. Conclusions The HBV isolated from patients in immune clearance phase have higher variability than those isolated from patients in immune tolerance phase. The longer the infection persists and the more severe the disease is, the more variable HBV quasispecies are.

关 键 词：肝炎 乙型 慢性 肝硬化 肝炎病毒 突变 遗传异质性 

分 类 号：R51[医药卫生—内科学]