Long PCR and its application to hepatitis viruses: amplification of hepatitis A, hepatitis B, and hepatitis C virus genomes.

Journal of Clinical Microbiology
R TellierR H Purcell

Abstract

In this study we amplified virtually the entire genomes of hepatitis A virus (a member of the Picornaviridae family), hepatitis B virus (a member of the Hepadnaviridae family), and hepatitis C virus (a member of the Flaviviridae family) by using the recently described technique of long PCR. In order to do this, we first demonstrated, using the lambda phage, that long PCR can be made highly sensitive and the sensitivity can be further enhanced by nested long PCR. We also showed, using tobacco mosaic virus as a model, that a reverse transcriptase reaction can be linked to a long PCR, enabling the nearly full-length amplification of the genomes of RNA viruses. We then applied these techniques to serial dilutions of titrated stocks of well-characterized strains of hepatitis A, B, and C viruses. We amplified the nearly full-length sequence of each of these viruses from a small number of viral genomes, demonstrating the sensitivity of the process.

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Citations

Dec 26, 2001·The Journal of General Virology·Karen Z ChingBetty H Robertson
Feb 12, 2005·Acta Microbiologica Et Immunologica Hungarica·A PárB Hunyady
May 27, 2014·Food and Environmental Virology·Chiara ChiapponiStefano Pongolini
Feb 26, 2016·Viruses·Jani Ylä-PeltoPetri Susi
Feb 12, 1998·Proceedings of the National Academy of Sciences of the United States of America·X FornsS U Emerson
Jun 3, 2005·AIDS Research and Human Retroviruses·Vera HolzmayerJohn Hackett
Jul 27, 2007·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Jordan J FeldE Jenny Heathcote
Aug 5, 1997·Proceedings of the National Academy of Sciences of the United States of America·M YanagiJ Bukh
Sep 21, 2005·Journal of Medical Virology·Shintaro YagiKenjiro Yamaguchi
Jul 9, 1997·Biochemical and Biophysical Research Communications·R W ChamberlainR M Elliott
Aug 21, 2003·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·Eleni KostourosRaymond Tellier

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