Real‐time NASBA detection of SARS‐associated coronavirus and comparison with real‐time reverse transcription‐PCR
Identifieur interne : 000394 ( Istex/Corpus ); précédent : 000393; suivant : 000395Real‐time NASBA detection of SARS‐associated coronavirus and comparison with real‐time reverse transcription‐PCR
Auteurs : Maria Cristina Keightley ; Peter Sillekens ; Wim Schippers ; Charles Rinaldo ; Kirsten St. GeorgeSource :
- Journal of Medical Virology [ 0146-6615 ] ; 2005-11.
English descriptors
- Teeft :
- Assay, Chan, Clin, Clin microbiol, Clinical presentation, Coronavirus, Dean erdman, Different combinations, Dilution series, Disease control, Disease onset, Drosten, Early detection, Early diagnosis, Emery, European quality assurance, Genome, Greater sensitivity, Greatest sensitivity, High titer, Hong kong, Leader sequence, Lower case, Microbiol, Molecular beacons, Molecular tests, Nasba, Nasba application, Nasba assay, Nasba detection, Novel coronavirus, Nucleic, Nucleic acid, Peiris, Polymerase, Polymerase gene, Primer, Primer sets, Respiratory syndrome, Respiratory syndrome coronavirus, Respiratory syndrome diagnosis, Sars, Sars coronavirus, Sars patients, Syndrome, Taqman, Taqman sars, Taqman test, Urbani, Urbani strain, Viral, Viral load, Whole genome urbani strain, World health organization multicentre.
Abstract
Severe acute respiratory syndrome (SARS) exhibits a high mortality rate and the potential for rapid epidemic spread. Additionally, it has a poorly defined clinical presentation, and no known treatment or prevention methods. Collectively, these factors underscore the need for early diagnosis. Molecular tests have been developed to detect SARS coronavirus (SARS‐CoV) RNA using real time reverse transcription polymerase chain reaction (RT‐PCR) with varying levels of sensitivity. However, RNA amplification methods have been demonstrated to be more sensitive for the detection of some RNA viruses. We therefore developed a real‐time nucleic acid sequence‐based amplification (NASBA) test for SARS‐CoV. A number of primer/beacon sets were designed to target different regions of the SARS‐CoV genome, and were tested for sensitivity and specificity. The performance of the assays was compared with RT‐PCR assays. A multi‐target real‐time NASBA application was developed for detection of SARS‐CoV polymerase (Pol) and nucleocapsid (N) genes. The N targets were found to be consistently more sensitive than the Pol targets, and the real‐time NASBA assay demonstrates equivalent sensitivity when compared to testing by real‐time RT‐PCR. A multi‐target real‐time NASBA assay has been successfully developed for the sensitive detection of SARS‐CoV. J. Med. Virol. 77:602–608, 2005. © 2005 Wiley‐Liss, inc.
Url:
DOI: 10.1002/jmv.20498
Links to Exploration step
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<abstract lang="en">Severe acute respiratory syndrome (SARS) exhibits a high mortality rate and the potential for rapid epidemic spread. Additionally, it has a poorly defined clinical presentation, and no known treatment or prevention methods. Collectively, these factors underscore the need for early diagnosis. Molecular tests have been developed to detect SARS coronavirus (SARS‐CoV) RNA using real time reverse transcription polymerase chain reaction (RT‐PCR) with varying levels of sensitivity. However, RNA amplification methods have been demonstrated to be more sensitive for the detection of some RNA viruses. We therefore developed a real‐time nucleic acid sequence‐based amplification (NASBA) test for SARS‐CoV. A number of primer/beacon sets were designed to target different regions of the SARS‐CoV genome, and were tested for sensitivity and specificity. The performance of the assays was compared with RT‐PCR assays. A multi‐target real‐time NASBA application was developed for detection of SARS‐CoV polymerase (Pol) and nucleocapsid (N) genes. The N targets were found to be consistently more sensitive than the Pol targets, and the real‐time NASBA assay demonstrates equivalent sensitivity when compared to testing by real‐time RT‐PCR. A multi‐target real‐time NASBA assay has been successfully developed for the sensitive detection of SARS‐CoV. J. Med. Virol. 77:602–608, 2005. © 2005 Wiley‐Liss, inc.</abstract>
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