Establishment and application of real-time fluoriescene-based metry polymerase chain reaction for hepatitis B virus genotyping

Abstract

Abstract: Objective To explore a new method for determining hepatitis B virus (HBV) genotypes B-D with real-time fluorescence-based PCR. Methods The PCR primers and probes were designed according to the analysis of 143 complete HBV genomes from GenBank and on the basis of a search for genotype-specific nucleotide sequences which were conserved in the 3 HBV genotypes. Real-time fluorescence-based PCR was performed for HBV genotyping of 128 samples collected from Lanzhou. Three samples of each genotype of B, C and D were randomly selected and their S gene was sequenced for confirmation of the results of PCR-based method. Results Real-time PCR identified genotype B in 26 (20.3%), genotype C in 92 (71.9%), and genotype D in 10 (7.8%) cases. The sequencing results of the S gene of 18 PCR-produced clones were in complete consistence with those of fluorescence-based PCR. Conclusion The real-time PCR method is convenient, highly sensitive, rapid and accurate, especially suitable in clinical and large-scale epidemiological studies.

CLC Number:

SHEN Jian-kun¹, ZHOU Rong², WANG Zhan-hui¹, BAI Pei-sheng², MA Shi-wu¹, ZHANG Ke¹, HOU Jin-lin¹. Establishment and application of real-time fluoriescene-based metry polymerase chain reaction for hepatitis B virus genotyping[J]. Journal of Southern Medical University, 2005, 25(06): 630-632.

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