Study And Development Of Hepatitis B Virus Gene Mutation Detection And Analysis Platform

Hepatitis B virus (HBV) infection is a major health problem worldwide resulting inmultiple organ damages including cirrhosis, liver cancer and other chronic diseases. HBVgene has been identified in various genotypes, and exhibits highly mutagenic activity.Furthermore, the mutations of HBV play an important role in virus inducing drugresistance and chronic diseases. These indicate that the most basically work of theindividual diagnosis is to analyze the HBV mutation acquiring the associated informationof HBV genome in drug resistance and associated liver diseases. Until recently, theexisting sequence alignment algorithm is not suitable for HBV gene sequence analysis,because of its poor accuracy and applied limitation. Whereas, the non-computertechnology has a low efficiency in comparing gene mutations and analyzing genotypes.The most striking problem is the international gene analysis system is mainly based on thestandards and databases from foreign countries, which have some differences fromChinese population.Therefore, we studied and established the HBV Drug Guide analysis systemdepending on the molecular biology theories and the rapid development of bioinformatics.This system is based on NCBI database, combined with the Blast algorithm and Clustal algorithm to classify and analyze the mutation in P gene and BCP-PreC/C gene of HBV inthe application of personalized medicine direction. In this study, we found that HBV DrugGuide analysis system can be applied to detect the HBV mutation accurately in one time,direct the drug resistance, and determine the progression of HBV mutation and hepatoma.This analysis system provides more accurate, facility and important tool in in largequantities of HBV mutation detection and individual therapy combined with clinicalinformation.Experiment one: Design of HBV drug guideAccording to the theory and techniques of molecular biology and bioinformatics, weanalyze the feasibility and innovation of HBV drug guide system development.Furthermore, the associated bioinformatics theories and techniques were developedthrough bioinformatic analysis methods.Experiment two: Development of HBV drug guideIn this part, we analyzed the HBV Drug Guide system fully and each functionalmodule. Then we contructed the HBV Drug Guide to analyze from A to D of HBVgenotype based on the VC++. Net platform and modular programming ideas.Experiment three: Hepatitis B patients testing based on HBV drug guide softwarePeripheral blood serums of hepatitis B virus patients were collected. According to theP gene and PreC/C gene, the primers were designed and synthesized. PCR amplificationwas performed and sequenced to identify the HBV mutation of samples DNA. Accordingto the results analyzed by HBV Drug Guide System, we acquired and compared HBVmutation information from amount of samples, which including lots of variations can’t bedissolved by other software or systems. HBV Drug Guide System has shown its accuracyand practicality character comparing to Clustal X, Sequence scanner and et al. The resultsshowed that our system could be used to analyzed the gene sequence as normal, calculatethe QV value, and identify the mutation location exactly occurring in the database or a new mutation never been recognized. Moreover, this system also could be used to describethe drug resistance according to the HBV gene mutation, prognosis the progression ofHBV infection, and provide a powerful tool to support the mechanism research andtechnique direction in the further study of HBV infection. It is a useful platform in thedetection and analysis for hepatitis B virus gene mutation.