Real-time NASBA detection of SARS-associated coronavirus and comparison with real-time reverse transcription-PCR
Identifieur interne : 005111 ( Main/Merge ); précédent : 005110; suivant : 005112Real-time NASBA detection of SARS-associated coronavirus and comparison with real-time reverse transcription-PCR
Auteurs : Maria Cristina Keightley [États-Unis] ; Peter Sillekens [Pays-Bas] ; Wim Schippers [Pays-Bas] ; Charles Rinaldo [États-Unis] ; Kirsten St. George [États-Unis]Source :
- Journal of medical virology [ 0146-6615 ] ; 2005.
Descripteurs français
- Pascal (Inist)
English descriptors
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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.
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Pennsylvania</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ></inist:fA14><country>États-Unis</country><placeName><region type="state">Pennsylvanie</region></placeName></affiliation><affiliation wicri:level="4"><inist:fA14 i1="02"><s1>Department of Pathology, University of Pittsburgh</s1><s2>Pittsburgh, Pennsylvania</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ></inist:fA14><country>États-Unis</country><placeName><region type="state">Pennsylvanie</region><settlement type="city">Pittsburgh</settlement></placeName><orgName type="university">Université de Pittsburgh</orgName></affiliation></author><author><name sortKey="Sillekens, Peter" sort="Sillekens, Peter" uniqKey="Sillekens P" first="Peter" last="Sillekens">Peter Sillekens</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Department of bioMérieux</s1><s2>Boxtel</s2><s3>NLD</s3><sZ>2 aut.</sZ><sZ>3 aut.</sZ></inist:fA14><country>Pays-Bas</country><wicri:noRegion>Department of bioMérieux</wicri:noRegion></affiliation></author><author><name sortKey="Schippers, Wim" sort="Schippers, Wim" uniqKey="Schippers W" first="Wim" last="Schippers">Wim Schippers</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Department of bioMérieux</s1><s2>Boxtel</s2><s3>NLD</s3><sZ>2 aut.</sZ><sZ>3 aut.</sZ></inist:fA14><country>Pays-Bas</country><wicri:noRegion>Department of bioMérieux</wicri:noRegion></affiliation></author><author><name sortKey="Rinaldo, Charles" sort="Rinaldo, Charles" uniqKey="Rinaldo C" first="Charles" last="Rinaldo">Charles Rinaldo</name><affiliation wicri:level="2"><inist:fA14 i1="01"><s1>Clinical Virology Laboratory, University of Pittsburgh Medical Center</s1><s2>Pittsburgh, Pennsylvania</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ></inist:fA14><country>États-Unis</country><placeName><region type="state">Pennsylvanie</region></placeName></affiliation><affiliation wicri:level="4"><inist:fA14 i1="02"><s1>Department of Pathology, University of Pittsburgh</s1><s2>Pittsburgh, Pennsylvania</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ></inist:fA14><country>États-Unis</country><placeName><region type="state">Pennsylvanie</region><settlement type="city">Pittsburgh</settlement></placeName><orgName type="university">Université de Pittsburgh</orgName></affiliation></author><author><name sortKey="George, Kirsten St" sort="George, Kirsten St" uniqKey="George K" first="Kirsten St." last="George">Kirsten St. George</name><affiliation wicri:level="2"><inist:fA14 i1="01"><s1>Clinical Virology Laboratory, University of Pittsburgh Medical Center</s1><s2>Pittsburgh, Pennsylvania</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ></inist:fA14><country>États-Unis</country><placeName><region type="state">Pennsylvanie</region></placeName></affiliation><affiliation wicri:level="4"><inist:fA14 i1="02"><s1>Department of Pathology, University of Pittsburgh</s1><s2>Pittsburgh, Pennsylvania</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ></inist:fA14><country>États-Unis</country><placeName><region type="state">Pennsylvanie</region><settlement type="city">Pittsburgh</settlement></placeName><orgName type="university">Université de Pittsburgh</orgName></affiliation></author></analytic><series><title level="j" type="main">Journal of medical virology</title><title level="j" type="abbreviated">J. med. virol.</title><idno type="ISSN">0146-6615</idno><imprint><date when="2005">2005</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Journal of medical virology</title><title level="j" type="abbreviated">J. med. virol.</title><idno type="ISSN">0146-6615</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Detection</term><term>Real time</term><term>Reverse transcription polymerase chain reaction</term><term>Severe acute respiratory syndrome</term><term>Severe acute respiratory syndrome virus</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Virus syndrome respiratoire aigu sévère</term><term>Temps réel</term><term>Détection</term><term>Réaction chaîne polymérase RT</term><term>Syndrome respiratoire aigu sévère</term></keywords></textClass></profileDesc></teiHeader><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></front></TEI><affiliations><list><country><li>Pays-Bas</li><li>États-Unis</li></country><region><li>Pennsylvanie</li></region><settlement><li>Pittsburgh</li></settlement><orgName><li>Université de Pittsburgh</li></orgName></list><tree><country name="États-Unis"><region name="Pennsylvanie"><name sortKey="Keightley, Maria Cristina" sort="Keightley, Maria Cristina" uniqKey="Keightley M" first="Maria Cristina" last="Keightley">Maria Cristina Keightley</name></region><name sortKey="George, Kirsten St" sort="George, Kirsten St" uniqKey="George K" first="Kirsten St." last="George">Kirsten St. George</name><name sortKey="George, Kirsten St" sort="George, Kirsten St" uniqKey="George K" first="Kirsten St." last="George">Kirsten St. George</name><name sortKey="Keightley, Maria Cristina" sort="Keightley, Maria Cristina" uniqKey="Keightley M" first="Maria Cristina" last="Keightley">Maria Cristina Keightley</name><name sortKey="Rinaldo, Charles" sort="Rinaldo, Charles" uniqKey="Rinaldo C" first="Charles" last="Rinaldo">Charles Rinaldo</name><name sortKey="Rinaldo, Charles" sort="Rinaldo, Charles" uniqKey="Rinaldo C" first="Charles" last="Rinaldo">Charles Rinaldo</name></country><country name="Pays-Bas"><noRegion><name sortKey="Sillekens, Peter" sort="Sillekens, Peter" uniqKey="Sillekens P" first="Peter" last="Sillekens">Peter Sillekens</name></noRegion><name sortKey="Schippers, Wim" sort="Schippers, Wim" uniqKey="Schippers W" first="Wim" last="Schippers">Wim Schippers</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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