乙型肝炎病毒共价闭合环状DNA的检测与临床应用

目的 建立检测乙型肝炎病毒(HBV)共价闭合环状DNA(cccDNA)聚合酶链反应法,研究其临床应用价值.方法 根据HBV-cccDNA核苷酸序列特点设计PCR引物,应用PCR的方法检测急性和慢性乙型病毒性肝炎、肝硬化、拉米夫定治疗等病患者的血清、外周血细胞及肝组织cccDNA.结果 急性及慢性乙型病毒性肝炎、拉米夫定治疗等病患者血清的HBV cccDNA检出阳性率分别为15.6％、50.0％、21.2％；前两者外周血细胞检出阳性率分别为10.7％、19.6％；慢性乙型病毒性肝炎(含肝硬化)、拉米夫定治疗等病患者肝组织的HBV-cccDNA检出阳性率分别为91.1％、32.8％.结论 该方法操作简单、特异性强,可用于HBV-ccc DNA的检测,评价抗乙型肝炎病毒药物的疗效.

Detection of HBV cccDNA and its clinical application

OBJECTIVE To establish the detection of hepatitis B virus covalently closed circular DNA(HBV cccDNA) polymerase chain reaction method,and study its clinical value.METHODS Based on HBV cccDNA sequences,the designed PCR primers were used to detect HBVcccDNA in serum,peripheral blood cells and infected liver cells of the patients with acute and chronic hepatitis B,liver cirrhosis,and the treatment with lamivudine.RESULTS The positive rates of HBV cccDNA in the serum of the patients with acute and chronic hepatitis B,lamivudine therapy were 15.6%,30.9%,and 21.2%,respectively;the positive rates of the peripheral blood cells of the former two were 10.7% and 19.6%,respectively;the positive rates of the HBV cccDNA in the liver tissue of the patients with chronic hepatitis B(including cirrhosis) and the treatment with lamivudine were 91.1% and 32.8%,respectively.CONCLUSION The method is simple and specific,and can be used for the detection of HBV cccDNA as well as the evaluation of the efficacy of anti-HBV drugs.