Evaluation And Optimization Of Assay For Specific Quantitative Detection Of Hepatitis B Virus CccDNA And Its Application

Hepatitis B virus(HBV) covalently closed circular DNA(cccDNA) accumulates in the nucleus of infect cells. It originates from virion DNA and serves as templates for the synthesis of pregenomic RNA and viral mRNAs. HBV cccDNA is now denoted as a marker of replication of HBV and its level may be consistant with the activity of its replication. The quantitative assay of cccDNA is a critical tool in the study of HBV clearance, the mechanism of anti-HBV agents and the follow up of HBV infected patients, however, the particular structure and biological characteristic of cccDNA pose a large obstacle to high specificity. Many cccDNA specifically determin assays have been developed since 1990s', but few of them have been widely accepted. Nearly all of our knowledge of cccDNA come from the study of cell models or animal models such as duck, wood chuck, chimpanzee. Only a little data comes from the patients infected with hepatitis B virus, which need to be enriched. We assessed some specifical determination of cccDNA assays from the reported articles to select some better ones. Then we optimized these methods and integrated them a single assay to detcet the pripheral blood mononuclear cell(PBMC) of patients in different period of post-liver-transplantation and the samples of liver puncturation in patients with chronic hepatitis B.This research was composed of three parts: â… . The accessment and optimization of specifical determination of cccDNA assays. â…¡. Quantitative detection of hepatitis B virus cccDNA in peripheral blood mononuclear cell and liver tissue in patients with chronic hepatitis B. HI Detection hepatitis B virus in the peripheral blood mononuclear cells of post-liver-transplantation patients associated with hepatitis B.â…  The evaluation and optimization of specifically determination of cccDNA assaysObjectives To evaluate several methods for specific quantitative detection of...