Classification of hepatitis B virus genotypes by the PCR-Invader method with genotype-specific probes.
Hepatitis B virus is a worldwide public health problem. A simple and effective test to identify viral genotypes would greatly aid efforts to understand and control the spread of this disease. A serial invasive signal amplification reaction assay (PCR-Invader assay) was developed for distinguishing the known eight genotypes (A-H) and four subgenotypes (Aa, Ae, Ba, Bj) of hepatitis B virus (HBV). The preS/S and core regions were amplified by multiplex PCR and delivered to 12 wells containing genotype-specific Invader probes. By observing the fluorescence patterns in the wells, HBV sub/genotypes can be assigned. A total of 505 serum samples containing HBV/HBsAg in Japan was examined by PCR-Invader and compared the results with those from ELISA assays with monoclonal antibodies against epitopes on gene products of the preS2 region and with a genotype-specific probe assay (GSPA) based on the preS1 region. Genotypes determined by the PCR-Invader agreed with those of the ELISA method in 98.2% of cases and with the GSPA method in 97.1% of cases. Co-infection with two distinct genotypes was correctly identified by the PCR-Invader in four serum samples, as determined by GSPA. Thus, the PCR-Invader assay is a useful tool for detecting the 10 known HBV sub/genotypes.
Tadokoro
Kenichi
K
Division of Advanced Technology and Development, BML, Inc., 1361-1, Matoba, Kawagoe-shi, Saitama 350-1101, Japan. tado-k@bml.co.jp
Kobayashi
Mariko
M
Yamaguchi
Toshikazu
T
Suzuki
Fumitaka
F
Miyauchi
Saeko
S
Egashira
Toru
T
Kumada
Hiromitsu
H
D001483:Base Sequence; D004797:Enzyme-Linked Immunosorbent Assay; D005456:Fluorescent Dyes / Q000032:analysis; D005838:Genotype; D006509:Hepatitis B / Q000821:virology*; D006512:Hepatitis B Core Antigens / Q000235:genetics; D006514:Hepatitis B Surface Antigens / Q000235:genetics; D006515:Hepatitis B virus / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D015336:Molecular Probe Techniques; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods*; D044967:Serum / Q000821:virology
D005456:Fluorescent Dyes; D006512:Hepatitis B Core Antigens; D006514:Hepatitis B Surface Antigens; D015345:Oligonucleotide Probes