METHOD FOR ASSAYING REPLICATION OF HBV AND TESTING SUSCEPTIBILITY TO DRUGS

摘要

A method for measuring the replication capacity of HBV, e.g. HBV present in a biological sample, possibly in the presence of a pharmaceutical product, in particular an antiviral agent, the method comprising:(a) possibly extracting nucleic acids contained in the biological sample;(b) PCR amplification of HBV nucleic acids using at least two primer pairs to obtain at least two amplified HBV genomic fragments representing more-than-full-length HBV genome;(c) cloning the fragments obtained under (b) into a vector, so producing a vector containing more-than-full-length HBV DNA;(d) transfecting or transducing susceptible cells with the vector;(e) culturing the transfected or transduced cells in conditions allowing synthesis of HBV pgRNA from the cloned HBV DNA;(f) possibly treating the cultured cells with the pharmaceutical product, in particular antiviral agent; and(g) determination of the replication capacity of the HBV, possibly incidence of the pharmaceutical product, in particular antiviral agent, on viral gene expression and/or viral replication.