Chou W H, Lin S F, Sheu S H, Lu C F, Lin S Y, Wu J S
National Institute of Preventive Medicine, Department of Health, Taipei, Taiwan, Republic of China.
Jpn J Med Sci Biol. 1993 Aug;46(4):191-202. doi: 10.7883/yoken1952.46.191.
To understand what genotypes of hepatitis C virus (HCV) exist in Taiwan, we chose the non-structure 5 (NS5) region of the HCV genome for the target area of reverse transcription-polymerase chain reaction (RT-PCR) to detect HCV RNA from sera of hemodialysis patients. Of 39 serum samples which were positive for the HCV antibody among 87 samples from hemodialysis patients, 12 (antibody against HCV core protein, OD > 2) were examined by the RT-PCR. The plasmid pUC19 was used to clone HCV cDNAs in the NS5 region (401 bp) derived from 11 serum samples which were positive for HCV RNA. Sequence analyses of individual clone of these 11 amplified cDNA fragments were performed. Dr. Cha's classification (16) suggested that two genotypes of HCV were found in these serum samples; type II (2/18.2%) and type III (9/81.8%). Our study indicates also that NS5 is an adequate target region to differentiate HCV strains derived from different patients in the same hospital. The analysis of the amplified cDNA in the NS5 region of the HCV genome will therefore provide suitable information to perform a molecular epidemiological study on the transmission routes of this important virus infection.
为了解台湾地区丙型肝炎病毒（HCV）的基因型，我们选择HCV基因组的非结构5（NS5）区域作为逆转录聚合酶链反应（RT-PCR）的靶区域，以检测血液透析患者血清中的HCV RNA。在87份血液透析患者的样本中，有39份HCV抗体呈阳性，对其中12份（抗HCV核心蛋白抗体，OD>2）进行了RT-PCR检测。用质粒pUC19克隆了来自11份HCV RNA阳性血清样本的NS5区域（401 bp）的HCV cDNA。对这11个扩增的cDNA片段的单个克隆进行了序列分析。Cha博士的分类方法（16）表明，在这些血清样本中发现了两种HCV基因型；II型（2/18.2%）和III型（9/81.8%）。我们的研究还表明，NS5是区分同一医院不同患者来源的HCV毒株的合适靶区域。因此，对HCV基因组NS5区域扩增的cDNA进行分析，将为开展这一重要病毒感染传播途径的分子流行病学研究提供合适的信息。
