Detection of TTV DNA and analysis of partial TTV gene sequence in different population of Guizhou area / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the prevalence of transfusion-transmitted virus (TTV) infection and analyze partial nucleotide sequence of TTV isolated from Guizhou area.</p><p><b>METHODS</b>A nested polymerase chain reaction (nPCR) assay with two-set of primers deduced from the first read frame of TTV genome was established, the prevalence of TTV DNA were detected in 395 sera collected from different population in Guizhou area. The partial genes of TTV DNA fragments amplified from two liver disease patients and one hemodialysis patient were analysed and then compared with gene from Japan strain.</p><p><b>RESULTS</b>The positive rates of TTV DNA in 62 normal adults, 37 blood donors, 50 hemodialysis patients, 107 intravenous drug users (IVDUs), 139 patients with liver diseases were 6.45% (4/62), 8.10%(3/37), 26.00% (13/50), 25.23% (27/107) and 16.54% (23/139), respectively. The homology of TTV DNA sequence was from 99% to 100% within three Guizhou strains. As compared with published sequence, the nucleotide homology is 98% between all three Guizhou strains and Japan strain.</p><p><b>CONCLUSIONS</b>There are TT virus infection in Guizhou area, the infection rates were higher in hemodialysis patients and IVDUs. The nucleotide homology was high between Guizhou strain and Japan strain.</p>

The association of hepatitis B virus precore/basic core promoter mutations with genotype and progression of liver disease / 中华消化杂志

Objective To study association of hepatitis B virus(HBV) precore (pre c)/basic core promoter(BCP) mutations with the genotype or the progression of liver disease. Methods The serum samples from 148 patients with HBV-relative diseases were collected, including 31 asymptomatic carriers, 32 with chronic hepatitis B (CHB), 40 with liver cirrhosis(LC) and 45 with hepatocellular carcinoma(HCC). The genes covering HBV pre c and BCP were amplified by nested polymerase chain reaction (nPCR). The PCR products were subjected to direct sequencing and the mutations in pre c 1896 and BCP 1762/1764 were determined by sequence analysis. HBV genotypes were also detected in the sera by restriction fragment length polymorphism based on S-gene PCR products. Results Of 148 serum samples of HBV, 128 were successfully genotyped and sequenced. There were 60 genotype B and 68 genotype C. The mutation in pre c (A1896) was significantly higher in genotype B than in genotype C (48.3% vs 29.34%, P

Comparative study on detection methods of mutations in hepatitis B virus precore and basic core promoter / 中华检验医学杂志

0.05). The sequence of five clones from one serum which was identified precore mutation by mPCR-RFLP were all A1896 mutant strains.Another serum identified mixture infection by mPCR-RFLP , one clone was A1896 mutant strain and four were G1896 wild strains.The results of mPCR-RFLP were verified by cloning.Conclusions Comparison with sequencing, the mPCR-RFLP method is simple,accurate and can be used in large-scale surveys and clinical research.