Hepatitis C Virus Genome Cloned
Near Full-Length Hepatitis C Virus Genome Cloned from Clinical Samples
Gene Therapy Weekly - Sep. 14, 2006
Sept. 14, 2006 - (NewsRx.com) -- A study from the United States has reported that near full-length hepatitis C virus (HCV) genome can be cloned from clinical samples.
"Long RT-PCR (LRP) amplification of RNA templates is sometimes difficult compared to long PCR of DNA templates. Among RNA templates, HCV represents an excellent example to challenge the potential of LRP technology due to its extensive secondary structures and its difficulty to be readily cultured in vitro," wrote X.F. Fan and colleagues, St. Louis University.
"The only source for viral genome amplification is clinical samples in which HCV is usually present at low titers. We have created a comprehensive optimization protocol that allows robust amplification of a 9.1 kb fragment of HCV, followed by efficient cloning into a novel vector. Detailed analyses indicate the lack of potential LRP-mediated recombination and the preservation of viral diversity," the authors reported.
The researchers concluded, "Thus, our LRP protocol could be applied for the amplification of other difficult RNA templates and may facilitate RNA virus research such as linked viral mutations and reverse genetics."
Fan and colleagues published their study in Biochemical and Biophysical Research Communications (Efficient amplification and cloning of near full-length hepatitis C virus genome from clinical samples. Biochem Biophys Res Commun, 2006;346(4):1163-1172).
For more information, contact X.F. Fan, St. Louis University, School of Medicine, Division of Gastroenterology & Hepatology, Center Liver, Department of Internal Medicine, St. Louis, MO 63110, USA.
Publisher contact information for the journal Biochemical and Biophysical Research Communications is: Academic Press Inc. Elsevier Science, 525 B St., Ste. 1900, San Diego, CA 92101-4495, USA.
Gene Therapy Weekly - Sep. 14, 2006
Sept. 14, 2006 - (NewsRx.com) -- A study from the United States has reported that near full-length hepatitis C virus (HCV) genome can be cloned from clinical samples.
"Long RT-PCR (LRP) amplification of RNA templates is sometimes difficult compared to long PCR of DNA templates. Among RNA templates, HCV represents an excellent example to challenge the potential of LRP technology due to its extensive secondary structures and its difficulty to be readily cultured in vitro," wrote X.F. Fan and colleagues, St. Louis University.
"The only source for viral genome amplification is clinical samples in which HCV is usually present at low titers. We have created a comprehensive optimization protocol that allows robust amplification of a 9.1 kb fragment of HCV, followed by efficient cloning into a novel vector. Detailed analyses indicate the lack of potential LRP-mediated recombination and the preservation of viral diversity," the authors reported.
The researchers concluded, "Thus, our LRP protocol could be applied for the amplification of other difficult RNA templates and may facilitate RNA virus research such as linked viral mutations and reverse genetics."
Fan and colleagues published their study in Biochemical and Biophysical Research Communications (Efficient amplification and cloning of near full-length hepatitis C virus genome from clinical samples. Biochem Biophys Res Commun, 2006;346(4):1163-1172).
For more information, contact X.F. Fan, St. Louis University, School of Medicine, Division of Gastroenterology & Hepatology, Center Liver, Department of Internal Medicine, St. Louis, MO 63110, USA.
Publisher contact information for the journal Biochemical and Biophysical Research Communications is: Academic Press Inc. Elsevier Science, 525 B St., Ste. 1900, San Diego, CA 92101-4495, USA.
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