Establishment of a new HBV genotyping method with PCR-RBD and its application.
- Author:
Guang YANG
1
;
Jin-huan CUI
;
Shu CHEN
;
Jian-hua SI
;
Jia-ju TAN
;
Pei-yuan LI
;
Xin MA
Author Information
- Publication Type:Journal Article
- MeSH: DNA, Viral; genetics; Female; Genotype; Hepatitis B; virology; Hepatitis B virus; classification; genetics; Humans; Male; Oligonucleotide Array Sequence Analysis; methods
- From: Chinese Journal of Hepatology 2004;12(11):677-680
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVEUsing PCR-RDB to establish a new method for HBV genotyping, and to survey the distribution of HBV genotypes in the Foshan area.
METHODSBiotin-labeled primers for amplification of HBV region X (nt1550-1789) were used to amplify extracted HBV DNA. HBV was genotyped by hybridization of the PCR products with immobilized specific probes (genotype A to F) on C membrane. Color development was achieved by adding POD and TMB. A judgment was made according to color reactions. The reliability of this new method was verified by gene sequencing. 300 samples of HBV DNA-positive sera from the Foshan area were genotyped using this assay.
RESULTSOf the 300 sera genotyped by PCR-RBD, 147 (49.0%) cases were genotype B, 136 (45.3%) were genotype C, 1 (0.3%) genotype D, and 12 (4.0%) were mixtures of genotype B and C, and 4 (1.3%) were mixtures of genotype C and D. No genotype A, E or F were found. The results of PCR-RDB genotyping were consistent with the results obtained with sequence analysis.
CONCLUSIONThis newly established HBV genotyping system proved to be sensitive, specific, precise and economic, and should be suitable for clinical practice and epidemic study. The results of HBV genotyping show that genotype B and C are the predominant genotypes in the Foshan area.
