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- Title
Development of a molecular-beacon-based multi-allelic real-time RT-PCR assay for the detection of human coronavirus causing severe acute respiratory syndrome (SARS-CoV): a general methodology for detecting rapidly mutating viruses.
- Authors
Hadjinicolaou, Andreas; Farcas, Gabriella; Demetriou, Victoria; Mazzulli, Tony; Poutanen, Susan; Willey, Barbara; Low, Donald; Butany, Jagdish; Asa, Sylvia; Kain, Kevin; Kostrikis, Leondios
- Abstract
Emerging infectious diseases have caused a global effort for development of fast and accurate detection techniques. The rapidly mutating nature of viruses presents a major difficulty, highlighting the need for specific detection of genetically diverse strains. One such infectious agent is SARS-associated coronavirus (SARS-CoV), which emerged in 2003. This study aimed to develop a real-time RT-PCR detection assay specific for SARS-CoV, taking into account its intrinsic polymorphic nature due to genetic drift and recombination and the possibility of continuous and multiple introductions of genetically non-identical strains into the human population, by using mismatch-tolerant molecular beacons designed to specifically detect the SARS-CoV S, E, M and N genes. These were applied in simple, reproducible duplex and multiplex real-time PCR assays on 25 post-mortem samples and constructed RNA controls, and they demonstrated high target detection ability and specificity. This assay can readily be adapted for detection of other emerging and rapidly mutating pathogens.
- Subjects
SARS disease; CORONAVIRUSES; DETECTION of microorganisms; REVERSE transcriptase polymerase chain reaction; BIOLOGICAL assay; GENETIC recombination; VIRAL genetics
- Publication
Archives of Virology, 2011, Vol 156, Issue 4, p671
- ISSN
0304-8608
- Publication type
Article
- DOI
10.1007/s00705-010-0906-7