tailieunhanh - báo cáo hóa học:" A general method for nested RT-PCR amplification and sequencing the complete HCV genotype 1 open reading frame"

Tuyển tập các báo cáo nghiên cứu về hóa học được đăng trên tạp chí sinh học đề tài : A general method for nested RT-PCR amplification and sequencing the complete HCV genotype 1 open reading frame | Virology Journal BioMed Central Methodology Open Access A general method for nested RT-PCR amplification and sequencing the complete HCV genotype 1 open reading frame Ermei Yao1 John E Tavis 1 2 and the Virahep-C Study Group Address department of Molecular Microbiology and Immunology Saint Louis University School of Medicine Saint Louis Missouri 63104 USA and 2Saint Louis University Liver Center Saint Louis University School of Medicine Saint Louis Missouri 63104 USA Email Ermei Yao - yaoe@ John E Tavis - tavisje@ the Virahep-C Study Group - tavisje@ Corresponding author Published 01 December 2005 Received 17 June 2005 Accepted 01 December 2005 Virology Journal 2005 2 88 doi 1743-422X-2-88 This article is available from http content 2 1 88 2005 Yao et al licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License http licenses by which permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited. Abstract Background Hepatitis C virus HCV is a pathogenic hepatic flavivirus with a single stranded RNA genome. It has a high genetic variability and is classified into six major genotypes. Genotype 1a and 1b cause the majority of infections in the USA. Viral genomic sequence information is needed to correlate viral variation with pathology or response to therapy. However reverse transcription-polymerase chain reaction RT-PCR of the HCV genome must overcome low template concentration and high target sequence diversity. Amplification conditions must hence have both high sensitivity and specificity yet recognize a heterogeneous target population to permit general amplification with minimal bias. This places divergent demands of the amplification conditions that can be very difficult to reconcile. Results RT and nested PCR conditions were optimized independently and systematically .