[Establishment of a new HBV genotyping method with PCR-RBD and its application].
OBJECTIVE
Using PCR-RDB to establish a new method for HBV genotyping, and to survey the distribution of HBV genotypes in the Foshan area.


METHODS
Biotin-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.


RESULTS
Of 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.


CONCLUSIONS
This 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.
Yang
Guang
G
Institute of Clinical/Medical Research, the First People's Hospital of Foshan, Foshan, Guangdong 528000, China.
Cui
Jin-huan
JH
Chen
Shu
S
Si
Jian-hua
JH
Tan
Jia-ju
JJ
Li
Pei-yuan
PY
Ma
Xin
X
D004279:DNA, Viral / Q000235:genetics*; D005260:Female; D005838:Genotype; D006509:Hepatitis B / Q000821:virology; D006515:Hepatitis B virus / Q000145:classification / Q000235:genetics*; D006801:Humans; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*
D004279:DNA, Viral