慢性乙型肝炎和肝细胞癌患者乙肝病毒BCP区1762/1764双突变的定量检测及其意义  

Detection and significance of double mutation in basic core promoter in hepatitis B virus in chronic hepatitis B and hepatocellular carcinoma

作　　者：申悦平 徐君 满晓波[2] 张园海 

机构地区：[1]镇江市第三人民医院感染科,江苏镇江212021 [2]第二军医大学东方肝胆外科医院门诊部,上海210438

出　　处：《江苏大学学报（医学版）》2011年第6期514-518,共5页Journal of Jiangsu University：Medicine Edition

基　　金：江苏省自然科学基金资助项目(BK2006083)

摘　　要：目的:观察乙型肝炎病毒基本核心启动子(basic core promoter,BCP)区1762/1764双突变和肝细胞癌发病间的关系及意义。方法:利用已构建的乙肝病毒BCP区A1762T/G1764A双突变复制质粒,设计特异性TaqManMGB探针,采用特异性探针实时荧光定量PCR(PSRT-PCR)方法对从未进行过抗病毒治疗的80例慢性乙肝患者和50例已发生肝癌的慢性乙肝患者患者分别予以血清和肝组织双突变定量检测,比较双突变在这两类患者血清和肝组织中的发生情况。结果:慢性乙型肝炎患者组血清突变率为41.25%(33/80),肝组织突变率为45.00%(36/80);肝细胞癌组血清突变率为68.00%(34/50),肝组织突变率为78.00%(39/50),两组间的差异有统计学意义(P<0.005)。每组各自血清突变率和肝组织突变率无明显差异。乙肝组中高病毒载量患者(HBV DNA≥106/ml,43例)血清和肝组织的突变率分别为58.14%(25/43)、62.79%(27/43),低病毒载量患者(HBV DNA<106/ml,37例)血清和肝组织突变率分别为21.62%(8/37)、24.32%(9/37)。高病毒载量患者血清及肝组织双突变率明显高于低病毒载量组(P均<0.01)。结论:已发生肝癌的慢性乙肝患者BCP区双突变发生率明显高于未发生肝癌者,提示此双突变和肝癌的发生密切相关,检测此双突变有助于预测肝癌的发生;且血清检测和肝组织检测具有良好一致性。未发生肝癌的慢性乙肝患者病毒载量高提示双突变发生率高,及早进行抗病毒等治疗有助于降低双突变发生率。Objective： To explore the feasibility of quantitative detection of double mutation in basic core promoter（BCP） 1762/1764 sites in hepatitis B virus（HBV） in chronic hepatitis B（CHB） and hepatocellular carcinoma（HCC） and the relationship between the mutation and HCC and its clinical significance.Methods： The mutant was accurately detected using probe-specific real-time quantitative polymerase chain reaction（PSRT-PCR） which adopted TaqMan MGB probe,the designing was depended on the double mutation plasmids which were constructed based on the wild recombinant replication competent plasmids of HBV.Their different effluence was compared between CHB and HCC.Results： After detecting the serum and hepatic tissue in 80 CHB patients and 50 HCC patients（all patients never accepted antiviral treatment）,the HCC group expressed a higher mutant rate（68.00% vs 41.25%,P0.005） in the serum,as well as in the hepatic tissue（78.00% vs 45.00%,P0.005） respectively.The difference of mutation rate between the serum and the hepatic tissue was not significant either in CHB group and HCC group.In 80 CHB patients,compared with low viral load group（HBV DNA 106 copies/ml,37 cases）,high viral load group（HBV DNA≥106 copies/ml,43 cases） expressed a higher mutant rate not only in the serum（58.14% vs 21.62%,P0.005）,but also in the hepatic tissue（62.79% vs 24.32%,P0.005）.Conclusion： The higher rate of double mutation in patients with HCC hinted that the double mutation was related to the occurrence of HCC,which can be forecasted by detecting the double mutation.Based on the satisfactory consistency in detecting serum and hepatic tissue,to detect the patients′serums who could not accept drawing the tissue from their liver can accurately reflect their states of double mutation too,and it can easily be wide spreaded in clinic.High viral load hinted high mutation rate in hepatitis B patients without HCC,so furthermore early antiviral treatment can help to decrease the mutation rate.

关 键 词：肝炎病毒 基本核心启动子 基因突变 乙型肝炎 肝细胞癌 

分 类 号：R446.5[医药卫生—诊断学] R512.61[医药卫生—临床医学]