Real-time NASBA detection of SARS-associated coronavirus and comparison with real-time reverse transcription-PCR.
Identifieur interne : 001235 ( Ncbi/Merge ); précédent : 001234; suivant : 001236Real-time NASBA detection of SARS-associated coronavirus and comparison with real-time reverse transcription-PCR.
Auteurs : Maria Cristina Keightley [États-Unis] ; Peter Sillekens ; Wim Schippers ; Charles Rinaldo ; Kirsten St GeorgeSource :
- Journal of medical virology [ 0146-6615 ] ; 2005.
Descripteurs français
- KwdFr :
- MESH :
- analyse : ARN viral.
- diagnostic : Syndrome respiratoire aigu sévère.
- isolement et purification : Virus du SRAS.
- virologie : Syndrome respiratoire aigu sévère.
- Humains, RT-PCR, Techniques d'amplification d'acides nucléiques.
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
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pubmed:16254971Le document en format XML
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uniqKey="Schippers W" first="Wim" last="Schippers">Wim Schippers</name></author><author><name sortKey="Rinaldo, Charles" sort="Rinaldo, Charles" uniqKey="Rinaldo C" first="Charles" last="Rinaldo">Charles Rinaldo</name></author><author><name sortKey="George, Kirsten St" sort="George, Kirsten St" uniqKey="George K" first="Kirsten St" last="George">Kirsten St George</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2005">2005</date><idno type="RBID">pubmed:16254971</idno><idno type="pmid">16254971</idno><idno type="doi">10.1002/jmv.20498</idno><idno type="wicri:Area/PubMed/Corpus">002473</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002473</idno><idno type="wicri:Area/PubMed/Curation">002473</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">002473</idno><idno type="wicri:Area/PubMed/Checkpoint">002547</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" 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Sillekens</name></author><author><name sortKey="Schippers, Wim" sort="Schippers, Wim" uniqKey="Schippers W" first="Wim" last="Schippers">Wim Schippers</name></author><author><name sortKey="Rinaldo, Charles" sort="Rinaldo, Charles" uniqKey="Rinaldo C" first="Charles" last="Rinaldo">Charles Rinaldo</name></author><author><name sortKey="George, Kirsten St" sort="George, Kirsten St" uniqKey="George K" first="Kirsten St" last="George">Kirsten St George</name></author></analytic><series><title level="j">Journal of medical virology</title><idno type="ISSN">0146-6615</idno><imprint><date when="2005" type="published">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Humans</term><term>Nucleic Acid Amplification Techniques (methods)</term><term>RNA, Viral (analysis)</term><term>Reverse Transcriptase Polymerase Chain Reaction</term><term>SARS Virus (isolation & purification)</term><term>Severe Acute Respiratory Syndrome (diagnosis)</term><term>Severe Acute Respiratory Syndrome (virology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN viral (analyse)</term><term>Humains</term><term>RT-PCR</term><term>Syndrome respiratoire aigu sévère (diagnostic)</term><term>Syndrome respiratoire aigu sévère (virologie)</term><term>Techniques d'amplification d'acides nucléiques ()</term><term>Virus du SRAS (isolement et purification)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>RNA, Viral</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>ARN viral</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Nucleic Acid Amplification Techniques</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Humans</term><term>Reverse Transcriptase Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Humains</term><term>RT-PCR</term><term>Techniques d'amplification d'acides nucléiques</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><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">16254971</PMID><DateCompleted><Year>2006</Year><Month>01</Month><Day>17</Day></DateCompleted><DateRevised><Year>2006</Year><Month>11</Month><Day>15</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0146-6615</ISSN><JournalIssue CitedMedium="Print"><Volume>77</Volume><Issue>4</Issue><PubDate><Year>2005</Year><Month>Dec</Month></PubDate></JournalIssue><Title>Journal of medical virology</Title><ISOAbbreviation>J. Med. Virol.</ISOAbbreviation></Journal><ArticleTitle>Real-time NASBA detection of SARS-associated coronavirus and comparison with real-time reverse transcription-PCR.</ArticleTitle><Pagination><MedlinePgn>602-8</MedlinePgn></Pagination><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. 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