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Báo cáo hóa học: " Virus detection and identification using random multiplex (RT)-PCR with 3'-locked random primers"

Tham khảo tài liệu 'báo cáo hóa học: " virus detection and identification using random multiplex (rt)-pcr with 3'-locked random primers"', luận văn - báo cáo phục vụ nhu cầu học tập, nghiên cứu và làm việc hiệu quả | Virology Journal BioMed Central Research Virus detection and identification using random multiplex RT -PCR with 3 -locked random primers Amy L Clem Jonathan Sims Sucheta Telang John W Eaton and Jason Chesney Open Access Address Molecular Targets Program Medical Oncology J.G. Brown Cancer Center University of Louisville Kentucky USA Email Amy L Clem - alclem01@gwise.louisville.edu Jonathan Sims - jonathan.t.sims@gmail.com Sucheta Telang - sucheta.telang@louisville.edu John W Eaton - eatonredox@aol.com Jason Chesney - jason.chesney@louisville.edu Corresponding author Published 28 June 2007 Received 27 April 2007 Accepted 28 June 2007 Virology Journal 2007 4 65 doi l0.ll86 l 743-422X-4-65 This article is available from http www.virologyj.cOm content 4 1 65 2007 Clem et al licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License http creativecommons.org licenses by 2.0 which permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited. Abstract Background PCR-based detection and identification of viruses assumes a known relatively stable genome. Unfortunately high mutation rates may lead to extensive changes in viral nucleic acid sequences making dedicated PCR primer use problematic. Furthermore in bioterrorism viral consensus sequences can be genetically modified as a countermeasure to RT-PCR and DNA chip detection. Accordingly there is a great need for the development of rapid and universal virus detection and identification technologies. Results We report herein that viral genomic DNA or RNA can be separated from host nucleic acids in plasma by filtration and nuclease digestion and randomly amplified in a single PCR using a mixture of primers designed to be resistant to primer-dimer amplification 5 -VVVVVVVVAA-3 V A G or C 38 or 6561 primers . We have termed this novel PCR method Random Multiplex RT -PCR since hundreds of overlapping PCR .