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Hepatitis B virus genotype assignment using restriction fragment length polymorphism patterns
Author(s) -
Mizokami Masashi,
Nakano Tatsunori,
Orito Etsuro,
Tanaka Yasuhito,
Sakugawa Hiroshi,
Mukaide Motokazu,
Robertson Betty H.
Publication year - 1999
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(99)00471-8
Subject(s) - genotyping , genotype , restriction enzyme , restriction fragment length polymorphism , restriction site , biology , hepatitis b virus , genetics , microbiology and biotechnology , restriction fragment , virology , gene , virus
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, Hph I, Nci I, Alw I, Ear I, and Nla IV. 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 Ear I in genotype B was absent in spite of its presence in all the other genotypes and genotype C has no restriction site for Alw I. Only genotype E is digested with Nci I, while only genotype F has a restriction site for Hph I. Genotype A can be distinguished by a single restriction enzyme site for Nla IV, 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.