The Study On Relation Between Genovariation Of HBV In Jiangxi Province And Clinic Of CHB And Effect Of Antiviral

PartⅠDetection for genotypes of hepatitis B virus in Jiangxi Province of China by nested PCR with multiplex pairs of primers and its clinical relevanceObjective:To determine the hepatitis B virus (HBV) genotypes in Jiangxi Province by nested PCR with multiplex pairs of genotype-specific primers.Methods:150 serum samples from HBV-DNA positive patients with chronic heptitis B virus infection were divided into 5 groups according to patient's diagnosis, with each group of 30 samples. The 5 groups were asymptomatic carrier, mild, moderate, severe hepatitis B and fulminant hepatitis B, respectively. Nested PCR with multiplex pairs of genotype- specific primers was used to determinate the genotypes of HBV.Results:Of the 150 patients, 118 (78.7%) were with genotype B, 31 (20.7%) with genotype C and 1(0.6%) with genotype D. The positive rates of HBV genotype C in patients with asymptomatic carrier, mild, moderate, severe hepatitis B and fulminant hepatitis B were 0%, 16.7%, 26.7%, 30.0% and 30.0%, respectively, which was significantly different (P<0.05) only between asymptomatic group and others. The HbeAg positivity in patients with genotype B and C infection were 83.9%(99/118)and 90.3%(28/31), respectively and the HbeAb positivity were 16.1%(19/118) and 9.7%(3/31), respectively(P>0.05).Conclusion: The predominant HBV genotypes in Jiangxi Province are confirmed to be genotypes B and C. HBV of genotype C causes more serious hepatitis than HBV of genotype B. PartⅡNested PCR-RFLP method of Jiangxi Province and hepatitis B virus (Ba and Bj) subtypes of the preliminary identificationObjective: To determinate the distribution of hepatitis B virus (HBV) subtypes (Ba and Bj) in Jiangxi Province, and to know the relation between HBV subgenotype and pathogenetic condition, in order to explain the mechanisms that HBV of same genotype can lead to different pathogenetic condition.Method: 60 cases of hepatitis B virus, which were identifyed to be B genotype by nested PCR with multiplex pairs of primers, are randomly selected, including 30 chronic HBV carriers and 30 chronic patients with hepatitis B virus. Nested PCR-RFLP was used to detect the subgenotype of 60 cases. Results: The subgenotypes of 60 cases of patients were all Ba subtype, and no one was Bj subtype.Conclusion: The B genotyped HBV may be mainly Ba subgenotype in Jiangxi province. The mechanisms that HBV of same genotype can lead to different pathogenetic condition may be related with other factors, such as quasispecies.PartⅢStudy on relevance of quasispecies of hepatitis B virus in Lamivudine-resistant by melt curveObjective: To study on the relation between quasispecies of hepatitis B virus and Lamivudine-resistant, and to guide clinical medication.Method: The randomly selected serum samples of the CHB patients who have been treating with Lamivudine are divided into 3 groups, including 30 cases of pretherapy, 30 cases of variation and 30 cases which have been virus rebound after stoped therapy. The HBV polymerase gene was amplificated by PCR, then melt curve was applied to analyze the quasispecies quantity of c/s gene of hepatitis B virus. Results: The quasispecies number of HBV polymerase gene before treatment was 8.37±0.93; the virus rebound group was 5.30±0.95, the virus variation group was 2.50±0.86. They compared with each other, P<0.05. The quasispecies number of HBV core gene before treatment was 6.33±1.64; the virus rebound group was 6.37±1.81, the virus variation group was 6.10±1.86. There were no significant difference among three groups (P>0.05). The quasispecies number of HBV S gene before treatment was 5.77±2.11; the virus rebound group was 6.17±1.93, the virus variation group was 5.23±1.85. They compared with each other,P>0.05.Conclusions: Quasispecies of hepatitis B virus polymerase gene are decreased gradually under the selection of lamivudine.The number of quasispecies increased while hepatits B virus varied, but quasispecies quantity was less than that before treatment. Compared with group of HBV P gene, both of the quasispecies of HBV core gene and that of Sgene have no significant changes in quantity.