Abstract：
In recent years, serum hepatitis B virus (HBV) RNA has been identified as a promising noninvasive surrogate biomarker of intrahepatic covalently closed circular DNA (cccDNA), detection of which requires an invasive liver biopsy in patients with chronic HBV infection. It is impractical to detect intrahepatic cccDNA as a routine diagnosis for chronic hepatitis B (CHB) patients in clinical management. Here, we describe a detailed protocol for serum HBV RNA quantification, which can reflect the activity of intrahepatic cccDNA. The procedure includes three major steps: (1) Simultaneous isolation of HBV DNA and RNA from patients\' serum, (2) DNase I digestion for removing HBV DNA contamination, and (3) HBV RNA quantification by one-step reverse transcription qPCR.
摘要：
近年来,血清乙型肝炎病毒（HBV）RNA已被确定为肝内共价闭合环状DNA（cccDNA）的有希望的非侵入性替代生物标志物，慢性HBV感染患者需要进行侵入性肝活检。在临床管理中检测肝内cccDNA作为慢性乙型肝炎（CHB）患者的常规诊断是不切实际的。这里,我们描述了血清HBVRNA定量的详细协议，可以反映肝内cccDNA的活性。该程序包括三个主要步骤：（1）从患者血清中同时分离HBVDNA和RNA，(2)DNaseⅠ消化去除HBVDNA沾染，和（3）通过一步逆转录qPCR定量HBVRNA。
