ScienceAsia 20 (1994): 115-124 |doi: 10.2306/scienceasia1513-1874.1994.20.115

SUBTYPING OF THAI HEPATITIS B VIRUS DNA BY POLYMERASE CHAIN REACTION

DUANTHANORM THAWARANANTHA,¹ KRUAVON BALACHANDRA,¹ JITTAPORN WATANASEREE,² CHUENCHIT BOONCHIRD,³ PAICHIT WARACHIT,¹ JAKKRISS BHUMISAWASDI¹ AND SOMSAK PANTUWATANA⁴

ABSTRACT: Polymerase chain reaction was modified and applied for subtyping of 200 sera of Thai hepatitis B virus(HBV) carriers. Four pairs of oligonucleotide primers which derived from certain amino acids coded by the S gene were utilized for subtyping. First 2 pairs which contained the variations at amino acid residues 68 and 122 of the S gene product were used to subtype for d and y determinants, while the latters which had the variations at residues 47 and 213 were used for r and w determinants. Southern-blot hybridization was used to confirm the amplified products as due to HBV-DNA. Furthermore, an immunologicalsubtyping with monoclonal antibodies (immunodiffusion) was also conducted to verify the PCR approach. The study revealed that all these Thai HBV-carriers were ad determinant positive which no ay were found at all, and they could be segregated as adr and adw subtypes as 81.0% and 19.0%, respectively.

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1 National Institute of Health, Department of Medical Sciences, Nonthaburi, 11000
2 Government of Pharmaceuticals Organization
3 Department of Biotechnology, Mahidol University
4 Department of Microbiology, Mahidol University, Bangkok, Thailand.

Received June 23, 1994