CAJ | 학술논문

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10例.收集患者外周血血清,抽提HBV DNA,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HBV RT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x2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组间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,LC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.
목적 연구HBV감염불동상태하혈청병독반전록매(RT)구준충특점화림상의의.방법 50례미접수항병독치료적HBV감염자분위만성HBV휴대자(ASC)조10례、만성을형간염(CHB)조30례화을형간염간경화(LC)조10례.수집환자외주혈혈청,추제HBV DNA,PCR확증RT구기인조후극륭、측서.매례환자획득15～30개서렬,진행HBV RT구적준충이질성분석급상관기인돌변통계등생물신식학분석.다균수비교채용방차분석,중위수비교채용비삼수검험분석,비계량자료비교채용x2검험.결과 총공측서1221개극륭,기중ASC조152개,CHB조780개,LC조289개.3조간기인형구성차이무통계학의의.준충복잡도위LC조＞CHB조＞ASC조,3조간차이유통계학의의(F=33.400,P＜0.05).준충리산도위LC조＞CHB조＞ASC조,LC조여CHB조급ASC조균차이유통계학의의(F=18.070,P＜0.05),CHB조여ASC조간차이무통계학의의.결론 만성HBV감염과정중,면역청제기비면역내수기구유경관적HBV변이보계,수착병정적연장화병정가중,HBV적준충추향복잡.
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 (152from ASC patients,780 from CHB patients and 289 from LC patients).Genotype distribution showed no difference among three groups.However,the quasispecies complexity showed significant differences 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.