荧光定量检测HBV cccDNA在慢性乙型肝炎患者中的应用  

The application of cccDNA fluorescence quantitative detection of hepatitis B virus in patients with chronic hepatitis B

作　　者：姚思敏[1] 袁静[1] 曾丽红[1] 吴驰[1] 戴炜[1] 王东[1] 

机构地区：[1]深圳市第三人民医院病毒性肝炎科,广东深圳518020

出　　处：《中西医结合肝病杂志》2010年第2期108-110,共3页Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases

基　　金：深圳市科技信息局课题(No.200802055)

摘　　要：目的:研究慢性乙型肝炎患者HBV共价闭合环状DNA(cccDNA)在外周血的分布、肝组织中含量及探讨抗病毒治疗对慢性乙型肝炎患者血清HBV cccDNA的影响。方法:随机选取HBV DNA阳性慢性乙型肝炎患者、肝硬化患者、重型肝炎患者共125例。使用实时荧光定量PCR法检测血清中cccDNA。选取36例慢性乙型肝炎患者肝组织和外周血样本,酶切后进行荧光定量PCR检测。60例慢性乙型肝炎患者,按1:1随机分配接受拉米夫定或干扰素治疗,所有患者均治疗24周以上;应用实时荧光定量聚合酶链反应技术检测慢性乙型肝炎患者0周、8周、12周、24周、HBV cccDNA及HBV DNA含量。结果:125例患者中cccDNA阳性率为71.2%,以肝硬化患者阳性率最低。肝硬化患者HBVcccDNA检出阳性率与慢性乙型肝炎患者及重型肝炎患者比较,差异有显著性意义(P<0.05)。肝组织中HBV cccDNA与肝组织总HBV DNA及血清HBV DNA存在相关性(P<0.05),HBeAg阳性组与阴性组患者比较差异有显著性意义(P<0.05)。患者接受抗病毒治疗后血清HBV DNA及HBV cccDNA下降。结论:乙型肝炎肝硬化患者外周血HBV cccDNA阳性率低,慢性乙型肝炎患者中HBeAg(+)组病毒复制较HBeAg(-)组活跃。抗病毒治疗对血清HBV cccDNA有抑制作用;血清HBV cccDNA可作为抗病毒治疗的重要监测指标。Objective：To study HBV covalently closed circular DNA （cccDNA） in the distribution of peripheral blood and liver tissue content in the patients of chronic HBV infections,investigate the impact of anti-viral treatment on serum HBV covalently closed circular DNA（cccDNA）of patients with chronic hepatitis B.Methods：Randomly selected a total of 125 cases with HBV DNA positive chronic hepatitis B,cirrhosis,severe hepatitis patients,using real-time fluorescence quantitative PCR for the detection of serum cccDNA.DNA were extracted from serum and liver biopsy samples of 36 patients with chronic hepatitis B,and were measured by the real-time fluorescence quantitative PCR assay.Sixty cases of chronic hepatitis B patients were randomly assigned to receive lamivudine or interferon treatment in a ratio of 1：1.Serum HBV cccDNA and HBV DNA levels were detected with real-time PCR at the time of 0,8,12,24 weeks.Results：The cccDNA-positive rate was 71.2% in 125 patients,to the lowest positive rate in patients with cirrhosis.The positive rate of HBV cccDNA had significant difference between patients with cirrhosis and chronic hepatitis B patients or severe hepatitis patients（P〈0.05）.There was a significant correlation between intra-hepatic cccDNA,intra-hepatic total HBV DNA and serum HBV DNA（P〈0.05）.Mean-while,the positive rate had significant difference between HBeAg-positive group and HBeAg-negative group（P〈0.05）.After receiving antiretroviral therapy,level of serum HBV DNA and HBV cccDNA was decreased.Conclusion：Positive rate of serum HBV cccDNA in patients with cirrhosis is low.Viral replication in HBeAg（＋） group is more active than in HBeAg（-） group.Anti-viral therapy has inhibition on serum HBV cccDNA;serum HBV cccDNA can be used as an important anti-viral treatment indicators for monitoring.

关 键 词：乙型肝炎病毒 CCCDNA 实时荧光定量PCR 抗病毒治疗 

分 类 号：R512.62[医药卫生—内科学]