Whole Nucleotide Sequence Analysis Of Chinese Hepatitis B Virus Genotype D,Establish And Preliminary Apply Of A New Genotyping Method Of HBV

ABSTRACT To explore the prevalence and whole nucleotide sequence characteristic of hepatitis B virus (HBV) D genotype in China, HBV genotypes were investigated in some areas of China using a genotyping method of based on the restriction fragment length polymorphism (RFLP) of an amplified segment of the HBV pre S region. Results suggested that genotype D was not less genotype in these areas. By the HBV full-length polynierase chain reaction (PCR) amplification and clone, the first Chinese HBV strain of genotype D complete nucleotide was sequenced and analyzed, GeneBank accession number is AF2808 17. The evolutional distance of this strain of genotype D is the closest to four Swedish strains of genotype D in total 30 wild strains of HBV genotype D of GeneBank, Asian is not predominance geographical region of genotype D distribution areas, the discrepancy of evolution of HBV strains of genotype D may result from the different origin of virus, migratory orientation of virus and the long choice of virus in the host of the different hereditary and heterogeneous immunity. 223 full-genomic sequences were genotyped with DNASIS program and nucleotide and amino acid sequences of S gene aligned with CLUSTALW program, Genotype specific regions were identified by the restriction enzymes. It should be useful for epidemiologic investigation and for analyzing the potential influence of genotype on the course of HBV infection because of its characteristics of high sensitive, differential and RFLP patterns recognized easy, the disadvantage of few published sequence, little type and complicated RFLP patterns were avoid. Direct sequencing S gene was done to confirm that HBV S gene PCR-RFLP genotyping method was correct. Two groups of 80 asymptomatic HBV carriers (AsC) and 120 liver cirrhosis (LC) patients in Gang Dong area of China were genotyped by this method. The results show that HBV genotypes of A, E and F were not found in this group of patients, genotype B and C are the major genotype in this area, genotype B is the main genotype in group of AsC, the distribution of HBV genotypes of same categories of liver diseases group is no statistically significant difference with the state of HBeAg, two groups of difference categories of liver diseases is a statistically significant difference in the distribution of HBV genotypes (p <0.01), genotype C showed an increasing trend from AsC to LC; In contrast, the distribution of genotype B and D showed a decreasing trend in the same order. These results suggested that genotype C is associated with the development of severe liver diseases, and genotype D has a relatively good prognosis.