Nakao T, Enomoto N, Takada N, Takada A, Date T
Division of Cancer Research, Kanazawa Medical University, Ishikawa, Japan.
J Gen Virol. 1991 Sep;72 ( Pt 9):2105-12. doi: 10.1099/0022-1317-72-9-2105.
Recently, we reported that hepatitis C virus (HCV) can be classified genetically into two types, HCV-K1 and HCV-K2, which show 67% and 71% identity at the nucleotide and amino acid sequence levels in a 340 bp region which encodes the NS5 gene Gly-Asp-Asp motif. To develop a rapid method to classify the genomes of HCV isolates, we identified restriction fragment length polymorphisms (RFLPs) in reverse transcriptase-polymerase chain reaction products encoding a portion of the NS5 gene. AluI and AccII enabled HCV to be classified into the K1 and K2 types, and Sau96I enabled classification into the K1 type, and the K2a and K2b subtypes. These RFLPs also generally allow Japanese isolates to be distinguished from the prototype (PT, an isolate from the U.S.A.), which is a K1 type. Sequence analysis of the 5'-untranslated regions of Japanese isolates revealed near identity between the K1 type and PT, and 93 to 94% identity between the K1 and K2 types, indicating that there are type K1- and K2-specific RFLPs in this region. Our results suggest that the nucleotide sequences of the K1 and K2 types are different throughout the HCV genome. The incidence of HCV types K1, K2a and K2b, and PT in 50 samples was 74%, 16%, 8% and 2%, respectively.
最近，我们报道丙型肝炎病毒（HCV）在基因上可分为两种类型，即HCV-K1和HCV-K2，在编码NS5基因甘氨酸-天冬氨酸-天冬氨酸基序的340bp区域，它们在核苷酸和氨基酸序列水平上的同一性分别为67%和71%。为开发一种快速分类HCV分离株基因组的方法，我们在编码NS5基因部分序列的逆转录-聚合酶链反应产物中鉴定出限制性片段长度多态性（RFLP）。AluI和AccII可将HCV分为K1和K2型，Sau96I可将其分为K1型以及K2a和K2b亚型。这些RFLP通常也能区分日本分离株与原型株（PT，一株来自美国的分离株），PT属于K1型。对日本分离株5'-非翻译区的序列分析显示，K1型与PT近乎相同，K1型和K2型之间的同一性为93%至94%，表明该区域存在K1型和K2型特异性RFLP。我们的结果表明，K1型和K2型的核苷酸序列在整个HCV基因组中均存在差异。在50个样本中，HCV K1型、K2a型、K2b型以及PT型的发生率分别为74%、16%、8%和2%。
