CAJ | 학술논문

目的探讨血清HBV全长基因组的准种特性及其与慢性乙型肝炎(CHB)重症化的关系.方法未接受过抗病毒治疗的CHB、慢性重型乙型肝炎(CSHB)各4例患者,扩增外周血清中HBV全长基因组并克隆、测序,每个标本选取14～16个克隆进行DNA测序及生物信息学分析.计量资料采用独立样本t检验,计数资料采用x2检验.结果 CHB组和CSHB组8例患者共获得120个HBV全长基因组序列.每例患者均为单一基因B型或C型感染.两组全长克隆序列中均发现G1896A突变、A1762T/G1764A、T1753C/G突变,以及前S2区、前S1区起始密码子缺失,CSHB组1例患者还发现A1762T/G1764A/C1766T突变,CSHB组发生一个或多个突变的克隆数占100.0%(60/60),明显多于CHB组的76.7%(46/60)(x2=15.85,P<0.01).在核苷酸和氨基酸水平上,CSHB组的全长基因组、S基因、X基因、P基因和反转录酶编码序列区的准种复杂度和平均遗传距离均比CHB组大,C基因的则比CHB组小,但差异均无统计学意义.结论慢性重型乙型肝炎患者血清中HBV的各种突变率及准种异质性差异均大于慢性乙型肝炎患者,可能与CHB重症化有关,值得扩大样本进一步研究其临床意义.
목적 탐토혈청HBV전장기인조적준충특성급기여만성을형간염(CHB)중증화적관계.방법 미접수과항병독치료적CHB、만성중형을형간염(CSHB)각4례환자,확증외주혈청중HBV전장기인조병극륭、측서,매개표본선취14～16개극륭진행DNA측서급생물신식학분석.계량자료채용독립양본t검험,계수자료채용x2검험.결과 CHB조화CSHB조8례환자공획득120개HBV전장기인조서렬.매례환자균위단일기인B형혹C형감염.량조전장극륭서렬중균발현G1896A돌변、A1762T/G1764A、T1753C/G돌변,이급전S2구、전S1구기시밀마자결실,CSHB조1례환자환발현A1762T/G1764A/C1766T돌변,CSHB조발생일개혹다개돌변적극륭수점100.0%(60/60),명현다우CHB조적76.7%(46/60)(x2=15.85,P<0.01).재핵감산화안기산수평상,CSHB조적전장기인조、S기인、X기인、P기인화반전록매편마서렬구적준충복잡도화평균유전거리균비CHB조대,C기인적칙비CHB조소,단차이균무통계학의의.결론 만성중형을형간염환자혈청중HBV적각충돌변솔급준충이질성차이균대우만성을형간염환자,가능여CHB중증화유관,치득확대양본진일보연구기림상의의.
Objective To characterize serum hepatitis B virus(HBV)full-length genome quasispecies and to investigate its ralationship with severe exacerbation of chronic hepatitis B(CHB).Methods HBV full-length genome was amplified and cloned from four treatment naive CHB patients and four treatment naive CSHB patients.Fourteen to sixteen clones per sample were selected,sequenced and analyzed by bioinformatics software.The measurement data was compared by independent-samples t test and count data was analyzed by x2 test. Results Totally 120 HBV fulllength genome sequences were obtained.All the patients had either genotype B or C virus monoinfection.One hundred percent clones(60/60)from CSHB patients showed mutations including G1896A,A1762T/G1764A(one patient even carried A1762T/G1764A/C1766T mutations),T1753C/G and start codon mutations in preS2,preS1,which were more common than those from CHB patients(46/60,76.7%;x2=15.85,P<0.01).The quasispecies complexity and diversity were higher in CSHB patients than CHB patients within full-length genome,S,X,P genes and reverse transcriptase region,but lower within C gene at both nucleotide and amino acid levels.But the difference were not statistically significant in all regions.Conclusion The mutation frequency and quasispeeies heterogeneity in HBV genome are higher in CSHB patients than in CHB patients,which may play a role in the severe exacerbation of CHB and needs further investigation in large scale studies.