Abstract
The hepatitis B virus (HBV) has been classified into eight genotypes (A-H) based on intergenotypic divergence of at least 8% in the complete nucleotide sequence or more than 4% in the S gene. To facilitate the investigation of the relationship between the efficacy of drug treatment and the mutation with specific genotype of HBV, we have established a new genotyping strategy based on a fragment of the HBV DNA polymerase gene. Pairwise sequence and phylogenetic analyses were performed using CLUSTAL V (DNASTAR) on the eight (A-H) standard full-length nucleotide sequences of HBV DNA from GenBank (NCBI) and the corresponding semi-nested PCR products from the HBV DNA polymerase gene. The differences in the semi-nested PCR fragments of the polymerase genes among genotypes A through F were greater than 4%, which is consistent with the intergenotypic divergence of at least 4% in HBV DNA S gene sequences. Genotyping using the semi-nested PCR products of the DNA polymerase genes revealed that only genotypes B, C, and D were present in the 50 cases, from Shenyang, China, with a distribution of 11 cases (22%), 25 cases (50%), and 14 cases (28%) respectively. These results demonstrate that our new genotyping method utilizing a fragment of the HBV DNA polymerase gene is valid and can be employed as a general genotyping strategy in areas with prevalent HBV genotypes A through F. In Shenyang, China, genotypes C, B, and D were identified with this new genotyping method, and genotype C was demonstrated to be the dominant genotype.
Highlights
Hepatitis B virus (HBV) infection is a substantial public health problem, with approximately 400 million virus carriers worldwide [1]
In order to facilitate the study of drug treatments, how lamivudine resistance develops from polymerase gene mutations [14,15], we established a new genotyping method using a fragment of the HBV
Most HBV patients screen positive for the HBV polymerase gene by semi-nested polymerase chain reaction (PCR) The purpose of the semi-nested PCR was to screen for polymerase gene-positive patients
Summary
Hepatitis B virus (HBV) infection is a substantial public health problem, with approximately 400 million virus carriers worldwide [1]. HBV has been classified into eight genotyped, designated as A-H, based on intergenotypic divergence of at least 8% in the complete nucleotide sequence or more than 4% in the S gene [2,3,4,5,6,7,8]. HBV genotypes have distinct geographical distributions and correlate with the severity of liver diseases. HBV genotype C is associated with more severe liver diseases than genotype B [9,10,11], and patients infected. In order to facilitate the study of drug treatments, how lamivudine resistance develops from polymerase gene mutations [14,15], we established a new genotyping method using a fragment of the HBV
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