丙型肝炎病毒基因组大片段的扩增与融合

Amplification and fusion of long fragments of hepatitis C virus genome.

作者信息

Wang L F, Radkowski M, Vargas H, Rakela J, Laskus T

机构信息

Division of Transplantation Medicine, Pittsburgh Transplantation Institute, University of Pittsburgh Medical Center, PA 15213, USA.

出版信息

J Virol Methods. 1997 Nov;68(2):217-23. doi: 10.1016/s0166-0934(97)00132-8.

DOI:10.1016/s0166-0934(97)00132-8

PMID:9389412

Abstract

The 'long PCR' was used for amplification of hepatitis C virus (HCV) subgenomic fragments from liver. After testing several commercially available systems, it was found that Tth as the major enzyme is superior to using Taq. Employing a mixture of Tth and Vent polymerase (rTth polymerase, XL, Perkin Elmer) it was possible to amplify 4.6-kb and 9-kb fragments from biological samples containing as little as 10(2) and 10(4) viral copies, respectively. It was also demonstrated that 'long PCR' is useful for joining together large size amplification products.

摘要

“长链PCR”用于从肝脏中扩增丙型肝炎病毒（HCV）亚基因组片段。在测试了几种市售系统后，发现以Tth作为主要酶比使用Taq更具优势。使用Tth和Vent聚合酶的混合物（rTth聚合酶，XL，珀金埃尔默公司），分别从含有低至10²和10⁴个病毒拷贝的生物样品中扩增出4.6 kb和9 kb的片段。还证明了“长链PCR”对于连接大尺寸扩增产物很有用。