Study And Clinical Application On Quantitative Detection To Hepatitis B Virus Mutation

Objective To study the conservative region of HBV YMDD motif and L528M mutation lead by the long-term lamivudine therapy could help to guide the treatment of Hepatitis B with Nucleoside analogues drugs. Meanwhile, to understand the relations 1762 (A-T) /1764 (G-A) double mutant in BCP region with severe hepatitis, cirrhosis and liver cancer. methods cloning related to the lamivudine - resistant of YMDD wild strain, YVDD mutant strain, YIDD mutant plasmid, 1762 (A-T) /1764 (G-A) double mutation in BCP Region of the wild type plasmid ,1762 single mutant plasmid, 1764 single mutant plasmids and 1762/1764 double mutant plasmid.A new probe based primers specific quantitative PCR method was used for accurate detection of mutations.Conclusion. 1.This plasmid as standard, the certification of the method can be used for both mutation detection. Use in the detection of YMDD and BCP mutation, test results show that this method was established successfully.2.The method used to detect the clinical lamivudine resistance YMDD mutation, found that the mutation mixed wild-type was indeed existed in 34 cases, 17.52%(34/194) of the total number of cases. Used the method could detect early the existence of mutant strains,guide the clinical treatment. 3.Used this method in the detection of BCP1762/1764 double mutant in hepatitis and liver cancer patients, found that in liver cancer patients mutation rate was higher than those hepatitis patients with no liver cancer.We thought that the double mutation in BCP was related to the occurrence of liver cancer.