Hepatitis B virus genotype assignment using restriction fragment length polymorphism patterns

FEBS Letters

Mizokami, Masashi³  Nakano, Tatsunori¹  Sakugawa, Hiroshi⁴  Robertson, Betty H.⁵  Mukaide, Motokazu²  Tanaka, Yasuhito¹  Orito, Etsuro¹ 

[1] Second Department of Medicine, Nagoya City University Medical School, Kawasumi, Mizuho, Nagoya 467-8601, Japan;Center of Molecular Biology and Cytogenetics, SRL, Inc., Tokyo, Japan;Department of Blood Transfusion, Nagoya City University Medical School, Nagoya, Japan;First Department of Medicine, University of the Ryukyus, Okinawa, Japan;Hepatitis Branch, Division of Viral and Rickettsial Diseases, Center for Disease Control and Prevention, Atlanta, GA, USA

关键词: Hepatitis B virus;    Genotyping;    Restriction fragment length polymorphism;    Restriction enzyme;    S gene;   

DOI  :  10.1016/S0014-5793(99)00471-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Hepatitis B virus (HBV) is classified into genotypes A–F, which is important for clinical and etiological investigations. To establish a simple genotyping method, 68 full-genomic sequences and 106 S gene sequences were analyzed by the molecular evolutionary method. HBV genotyping with the S gene sequence is consistent with genetic analysis using the full-genomic sequence. After alignment of the S sequences, genotype specific regions are identified and digested by the restriction enzymes, HphI, NciI, AlwI, EarI, and NlaIV. This HBV genotyping system using restriction fragment length polymorphism (RFLP) was confirmed to be correct when the PCR products of the S gene in 23 isolates collected from various countries were digested with this method. A restriction site for EarI in genotype B was absent in spite of its presence in all the other genotypes and genotype C has no restriction site for AlwI. Only genotype E is digested with NciI, while only genotype F has a restriction site for HphI. Genotype A can be distinguished by a single restriction enzyme site for NlaIV, while genotype D digestion with this enzyme results in two products that migrates at 265 and 186 bp. This simple and accurate HBV genotyping system using RFLP is considered to be useful for research on HBV.

【 授权许可】

Unknown