Real-time NASBA detection of SARS-associated coronavirus and comparison with real-time reverse transcription-PCR.
Identifieur interne : 002473 ( PubMed/Corpus ); précédent : 002472; suivant : 002474Real-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.
English descriptors
- KwdEn :
- MESH :
- chemical , analysis : RNA, Viral.
- diagnosis : Severe Acute Respiratory Syndrome.
- isolation & purification : SARS Virus.
- methods : Nucleic Acid Amplification Techniques.
- virology : Severe Acute Respiratory Syndrome.
- Humans, Reverse Transcriptase Polymerase Chain Reaction.
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.
DOI: 10.1002/jmv.20498
PubMed: 16254971
Links to Exploration step
pubmed:16254971Le document en format XML
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<author><name sortKey="Keightley, Maria Cristina" sort="Keightley, Maria Cristina" uniqKey="Keightley M" first="Maria Cristina" last="Keightley">Maria Cristina Keightley</name>
<affiliation><nlm:affiliation>Clinical Virology Laboratory, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania 15213, USA.</nlm:affiliation>
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<author><name sortKey="Sillekens, Peter" sort="Sillekens, Peter" uniqKey="Sillekens P" first="Peter" last="Sillekens">Peter Sillekens</name>
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<author><name sortKey="Schippers, Wim" sort="Schippers, Wim" uniqKey="Schippers W" first="Wim" last="Schippers">Wim Schippers</name>
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<author><name sortKey="Rinaldo, Charles" sort="Rinaldo, Charles" uniqKey="Rinaldo C" first="Charles" last="Rinaldo">Charles Rinaldo</name>
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<author><name sortKey="George, Kirsten St" sort="George, Kirsten St" uniqKey="George K" first="Kirsten St" last="George">Kirsten St George</name>
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<term>Reverse Transcriptase Polymerase Chain Reaction</term>
<term>SARS Virus (isolation & purification)</term>
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<front><div type="abstract" xml: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.</div>
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<Abstract><AbstractText>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.</AbstractText>
<CopyrightInformation>Copyright (c) 2005 Wiley-Liss, inc.</CopyrightInformation>
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