Improved multiplex-PCR to identify hepatitis B virus genotypes A-F and subgenotypes B1, B2, C1 and C2.
BACKGROUND
There are eight genotypes (A-H) and numerous subgenotypes of hepatitis B virus (HBV). The genotype has been shown to affect the course of HBV infection.


OBJECTIVE
To develop an efficient genotyping and subgenotyping method for large-scale epidemiological surveys of HBV infection in countries with high prevalence of HBV B and C such as China.


METHODS
We designed genotype and subgenotype-specific primer pairs, and adjusted PCR conditions for a multiplex-PCR using common Taq polymerase to identify HBV genotypes A-F in one reaction and for the main subgenotypes B1/B2 and C1/C2 in another reaction.


RESULTS
We have developed a multiplex-PCR system, which specifically amplifies DNA of HBV genotypes and the corresponding main subgenotypes B and C from the sera of HBV patients. Our patients were infected with HBV of subgenotypes B2 (n=18), C1 (n=2) and C2 (n=48). Eleven patients were doubly infected and three showed a triple infection with HBV A, B and C.


CONCLUSIONS
The low-cost multiplex-PCR for identification of HBV genotypes A-F and main subgenotypes of HBV B and C, is rapid, reliable and sufficient for large-scale epidemiological surveys and clinical studies.
Chen
Jinsong
J
Department of Epidemiology, Second Military Medical University, 800 Xiangyin Road, Shanghai 200433, PR China.
Yin
Jianhua
J
Tan
Xiaojie
X
Zhang
Haiqin
H
Zhang
Hongwei
H
Chen
Beichuan
B
Chang
Wenjun
W
Schaefer
Stephan
S
Cao
Guangwen
G
D001483:Base Sequence; D005838:Genotype; D006515:Hepatitis B virus / Q000145:classification / Q000235:genetics*; D019694:Hepatitis B, Chronic / Q000821:virology; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity