Rapid and sensitive detection of multiple genes from the SARS-coronavirus using quantitative RT-PCR with dual systems.
Identifieur interne : 001132 ( Ncbi/Merge ); précédent : 001131; suivant : 001133Rapid and sensitive detection of multiple genes from the SARS-coronavirus using quantitative RT-PCR with dual systems.
Auteurs : Jau-Ling Huang [Taïwan] ; Hui-Tsu Lin ; Yu-Ming Wang ; Yi-Chien Yeh ; Konan Peck ; Bai-Ling Lin ; Huan-Wun Liu ; Ann Chen ; Chang-Shen LinSource :
- Journal of medical virology [ 0146-6615 ] ; 2005.
Descripteurs français
- KwdFr :
- MESH :
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : Virus du SRAS.
- isolement et purification : Virus du SRAS.
- virologie : Syndrome respiratoire aigu sévère.
- ARN viral, Gènes viraux, Humains, RT-PCR, Sensibilité et spécificité.
English descriptors
- KwdEn :
- MESH :
- chemical : RNA, Viral.
- diagnosis : Severe Acute Respiratory Syndrome.
- genetics : SARS Virus.
- isolation & purification : SARS Virus.
- virology : Severe Acute Respiratory Syndrome.
- Genes, Viral, Humans, Reverse Transcriptase Polymerase Chain Reaction, Sensitivity and Specificity.
Abstract
The outbreak of severe acute respiratory syndrome (SARS) was caused by a newly identified coronavirus (SARS-CoV) in 2003. To detect early SARS-CoV infection, a one-step, real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) assay was developed that could simultaneously detect nucleocapsid (N), membrane (M), and spike (S) genes of SARS-CoV with the same PCR condition using either Applied Biosystems (ABI) Prism 7700 Sequence Detection System or Roche LightCycler. The sensitivity of this assay was evaluated using cell culture-derived viruses, in vitro transcribed viral RNA, and clinical specimens. The SARS-S, -M, and -N primer/probe sets described in this paper could detect one to ten copies of in vitro transcribed S, M, and N RNA per test using both the ABI and Roche assay systems. The relative sensitivities for detecting cell culture-derived SARS-CoV were 0.01, 0.01, and 0.001 PFU/test, respectively. It showed that SARS-N has comparable detection efficiencies to SARS2 and SARS3 which are primers sets designed by Centers for Disease Control and Prevention. In addition, SARS-S and SARS-M also demonstrated equivalent sensitivity to the commercially available RealArt HPA-Coronavirus reagents (Artus). The relative sensitivity of these primer/probe sets was also examined using human sera spiked viruses and clinical specimens from four confirmed SARS patients. Similar results as above were obtained. Specificity tests and sequence alignment showed that these primer/probe sets annealed perfectly to 31 isolates of SARS-CoV; and there was no cross detection with other coronaviruses and human respiratory tract-associated viruses. Therefore, not only is it compatible with the ABI and Roche systems, this multiple-gene detection assay also has the merit of being a rapid, safe, sensitive, and specific tool for accurate diagnosis of SARS-CoV infection.
DOI: 10.1002/jmv.20432
PubMed: 16121372
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JauLingH@mail.cju.edu.tw</nlm:affiliation><country xml:lang="fr">Taïwan</country><wicri:regionArea>Institute of Preventive Medicine, National Defense Medical Center, Taipei</wicri:regionArea><wicri:noRegion>Taipei</wicri:noRegion></affiliation></author><author><name sortKey="Lin, Hui Tsu" sort="Lin, Hui Tsu" uniqKey="Lin H" first="Hui-Tsu" last="Lin">Hui-Tsu Lin</name></author><author><name sortKey="Wang, Yu Ming" sort="Wang, Yu Ming" uniqKey="Wang Y" first="Yu-Ming" last="Wang">Yu-Ming Wang</name></author><author><name sortKey="Yeh, Yi Chien" sort="Yeh, Yi Chien" uniqKey="Yeh Y" first="Yi-Chien" last="Yeh">Yi-Chien Yeh</name></author><author><name sortKey="Peck, Konan" sort="Peck, Konan" uniqKey="Peck K" first="Konan" last="Peck">Konan Peck</name></author><author><name sortKey="Lin, Bai Ling" sort="Lin, Bai Ling" uniqKey="Lin B" first="Bai-Ling" last="Lin">Bai-Ling Lin</name></author><author><name sortKey="Liu, Huan Wun" sort="Liu, Huan Wun" uniqKey="Liu H" first="Huan-Wun" last="Liu">Huan-Wun Liu</name></author><author><name sortKey="Chen, Ann" sort="Chen, Ann" uniqKey="Chen A" first="Ann" last="Chen">Ann Chen</name></author><author><name sortKey="Lin, Chang Shen" sort="Lin, Chang Shen" uniqKey="Lin C" first="Chang-Shen" last="Lin">Chang-Shen Lin</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2005">2005</date><idno type="RBID">pubmed:16121372</idno><idno type="pmid">16121372</idno><idno type="doi">10.1002/jmv.20432</idno><idno type="wicri:Area/PubMed/Corpus">002571</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002571</idno><idno type="wicri:Area/PubMed/Curation">002571</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">002571</idno><idno type="wicri:Area/PubMed/Checkpoint">002553</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">002553</idno><idno type="wicri:Area/Ncbi/Merge">001132</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Rapid and sensitive detection of multiple genes from the SARS-coronavirus using quantitative RT-PCR with dual systems.</title><author><name sortKey="Huang, Jau Ling" sort="Huang, Jau Ling" uniqKey="Huang J" first="Jau-Ling" last="Huang">Jau-Ling Huang</name><affiliation wicri:level="1"><nlm:affiliation>Institute of Preventive Medicine, National Defense Medical Center, Taipei, Taiwan. JauLingH@mail.cju.edu.tw</nlm:affiliation><country xml:lang="fr">Taïwan</country><wicri:regionArea>Institute of Preventive Medicine, National Defense Medical Center, Taipei</wicri:regionArea><wicri:noRegion>Taipei</wicri:noRegion></affiliation></author><author><name sortKey="Lin, Hui Tsu" sort="Lin, Hui Tsu" uniqKey="Lin H" first="Hui-Tsu" last="Lin">Hui-Tsu Lin</name></author><author><name sortKey="Wang, Yu Ming" sort="Wang, Yu Ming" uniqKey="Wang Y" first="Yu-Ming" last="Wang">Yu-Ming Wang</name></author><author><name sortKey="Yeh, Yi Chien" sort="Yeh, Yi Chien" uniqKey="Yeh Y" first="Yi-Chien" last="Yeh">Yi-Chien Yeh</name></author><author><name sortKey="Peck, Konan" sort="Peck, Konan" uniqKey="Peck K" first="Konan" last="Peck">Konan Peck</name></author><author><name sortKey="Lin, Bai Ling" sort="Lin, Bai Ling" uniqKey="Lin B" first="Bai-Ling" last="Lin">Bai-Ling Lin</name></author><author><name sortKey="Liu, Huan Wun" sort="Liu, Huan Wun" uniqKey="Liu H" first="Huan-Wun" last="Liu">Huan-Wun Liu</name></author><author><name sortKey="Chen, Ann" sort="Chen, Ann" uniqKey="Chen A" first="Ann" last="Chen">Ann Chen</name></author><author><name sortKey="Lin, Chang Shen" sort="Lin, Chang Shen" uniqKey="Lin C" first="Chang-Shen" last="Lin">Chang-Shen Lin</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>Genes, Viral</term><term>Humans</term><term>RNA, Viral</term><term>Reverse Transcriptase Polymerase Chain Reaction</term><term>SARS Virus (genetics)</term><term>SARS Virus (isolation & purification)</term><term>Sensitivity and Specificity</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</term><term>Gènes viraux</term><term>Humains</term><term>RT-PCR</term><term>Sensibilité et spécificité</term><term>Syndrome respiratoire aigu sévère (diagnostic)</term><term>Syndrome respiratoire aigu sévère (virologie)</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (isolement et purification)</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>RNA, 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="genetics" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Virus du SRAS</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="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>Genes, Viral</term><term>Humans</term><term>Reverse Transcriptase Polymerase Chain Reaction</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>ARN viral</term><term>Gènes viraux</term><term>Humains</term><term>RT-PCR</term><term>Sensibilité et spécificité</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The outbreak of severe acute respiratory syndrome (SARS) was caused by a newly identified coronavirus (SARS-CoV) in 2003. To detect early SARS-CoV infection, a one-step, real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) assay was developed that could simultaneously detect nucleocapsid (N), membrane (M), and spike (S) genes of SARS-CoV with the same PCR condition using either Applied Biosystems (ABI) Prism 7700 Sequence Detection System or Roche LightCycler. The sensitivity of this assay was evaluated using cell culture-derived viruses, in vitro transcribed viral RNA, and clinical specimens. The SARS-S, -M, and -N primer/probe sets described in this paper could detect one to ten copies of in vitro transcribed S, M, and N RNA per test using both the ABI and Roche assay systems. The relative sensitivities for detecting cell culture-derived SARS-CoV were 0.01, 0.01, and 0.001 PFU/test, respectively. It showed that SARS-N has comparable detection efficiencies to SARS2 and SARS3 which are primers sets designed by Centers for Disease Control and Prevention. In addition, SARS-S and SARS-M also demonstrated equivalent sensitivity to the commercially available RealArt HPA-Coronavirus reagents (Artus). The relative sensitivity of these primer/probe sets was also examined using human sera spiked viruses and clinical specimens from four confirmed SARS patients. Similar results as above were obtained. Specificity tests and sequence alignment showed that these primer/probe sets annealed perfectly to 31 isolates of SARS-CoV; and there was no cross detection with other coronaviruses and human respiratory tract-associated viruses. Therefore, not only is it compatible with the ABI and Roche systems, this multiple-gene detection assay also has the merit of being a rapid, safe, sensitive, and specific tool for accurate diagnosis of SARS-CoV infection.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">16121372</PMID><DateCompleted><Year>2005</Year><Month>10</Month><Day>19</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>2</Issue><PubDate><Year>2005</Year><Month>Oct</Month></PubDate></JournalIssue><Title>Journal of medical virology</Title><ISOAbbreviation>J. Med. Virol.</ISOAbbreviation></Journal><ArticleTitle>Rapid and sensitive detection of multiple genes from the SARS-coronavirus using quantitative RT-PCR with dual systems.</ArticleTitle><Pagination><MedlinePgn>151-8</MedlinePgn></Pagination><Abstract><AbstractText>The outbreak of severe acute respiratory syndrome (SARS) was caused by a newly identified coronavirus (SARS-CoV) in 2003. To detect early SARS-CoV infection, a one-step, real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) assay was developed that could simultaneously detect nucleocapsid (N), membrane (M), and spike (S) genes of SARS-CoV with the same PCR condition using either Applied Biosystems (ABI) Prism 7700 Sequence Detection System or Roche LightCycler. The sensitivity of this assay was evaluated using cell culture-derived viruses, in vitro transcribed viral RNA, and clinical specimens. The SARS-S, -M, and -N primer/probe sets described in this paper could detect one to ten copies of in vitro transcribed S, M, and N RNA per test using both the ABI and Roche assay systems. The relative sensitivities for detecting cell culture-derived SARS-CoV were 0.01, 0.01, and 0.001 PFU/test, respectively. It showed that SARS-N has comparable detection efficiencies to SARS2 and SARS3 which are primers sets designed by Centers for Disease Control and Prevention. In addition, SARS-S and SARS-M also demonstrated equivalent sensitivity to the commercially available RealArt HPA-Coronavirus reagents (Artus). The relative sensitivity of these primer/probe sets was also examined using human sera spiked viruses and clinical specimens from four confirmed SARS patients. Similar results as above were obtained. Specificity tests and sequence alignment showed that these primer/probe sets annealed perfectly to 31 isolates of SARS-CoV; and there was no cross detection with other coronaviruses and human respiratory tract-associated viruses. Therefore, not only is it compatible with the ABI and Roche systems, this multiple-gene detection assay also has the merit of being a rapid, safe, sensitive, and specific tool for accurate diagnosis of SARS-CoV infection.</AbstractText><CopyrightInformation>(c) 2005 Wiley-Liss, Inc.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Huang</LastName><ForeName>Jau-Ling</ForeName><Initials>JL</Initials><AffiliationInfo><Affiliation>Institute of Preventive Medicine, National Defense Medical Center, Taipei, Taiwan. JauLingH@mail.cju.edu.tw</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Lin</LastName><ForeName>Hui-Tsu</ForeName><Initials>HT</Initials></Author><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Yu-Ming</ForeName><Initials>YM</Initials></Author><Author ValidYN="Y"><LastName>Yeh</LastName><ForeName>Yi-Chien</ForeName><Initials>YC</Initials></Author><Author ValidYN="Y"><LastName>Peck</LastName><ForeName>Konan</ForeName><Initials>K</Initials></Author><Author ValidYN="Y"><LastName>Lin</LastName><ForeName>Bai-Ling</ForeName><Initials>BL</Initials></Author><Author ValidYN="Y"><LastName>Liu</LastName><ForeName>Huan-Wun</ForeName><Initials>HW</Initials></Author><Author ValidYN="Y"><LastName>Chen</LastName><ForeName>Ann</ForeName><Initials>A</Initials></Author><Author ValidYN="Y"><LastName>Lin</LastName><ForeName>Chang-Shen</ForeName><Initials>CS</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>J Med Virol</MedlineTA><NlmUniqueID>7705876</NlmUniqueID><ISSNLinking>0146-6615</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D012367">RNA, Viral</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D005814" MajorTopicYN="Y">Genes, Viral</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D012367" MajorTopicYN="N">RNA, Viral</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D020133" MajorTopicYN="Y">Reverse Transcriptase Polymerase Chain Reaction</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D045473" MajorTopicYN="N">SARS Virus</DescriptorName><QualifierName UI="Q000235" MajorTopicYN="Y">genetics</QualifierName><QualifierName UI="Q000302" MajorTopicYN="N">isolation & purification</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D012680" MajorTopicYN="N">Sensitivity and Specificity</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D045169" MajorTopicYN="N">Severe Acute Respiratory Syndrome</DescriptorName><QualifierName UI="Q000175" MajorTopicYN="N">diagnosis</QualifierName><QualifierName UI="Q000821" MajorTopicYN="N">virology</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>2005</Year><Month>8</Month><Day>27</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2005</Year><Month>10</Month><Day>20</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2005</Year><Month>8</Month><Day>27</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">16121372</ArticleId><ArticleId IdType="doi">10.1002/jmv.20432</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Taïwan</li></country></list><tree><noCountry><name sortKey="Chen, Ann" sort="Chen, Ann" uniqKey="Chen A" first="Ann" last="Chen">Ann Chen</name><name sortKey="Lin, Bai Ling" sort="Lin, Bai Ling" uniqKey="Lin B" first="Bai-Ling" last="Lin">Bai-Ling Lin</name><name sortKey="Lin, Chang Shen" sort="Lin, Chang Shen" uniqKey="Lin C" first="Chang-Shen" last="Lin">Chang-Shen Lin</name><name sortKey="Lin, Hui Tsu" sort="Lin, Hui Tsu" uniqKey="Lin H" first="Hui-Tsu" last="Lin">Hui-Tsu Lin</name><name sortKey="Liu, Huan Wun" sort="Liu, Huan Wun" uniqKey="Liu H" first="Huan-Wun" last="Liu">Huan-Wun Liu</name><name sortKey="Peck, Konan" sort="Peck, Konan" uniqKey="Peck K" first="Konan" last="Peck">Konan Peck</name><name sortKey="Wang, Yu Ming" sort="Wang, Yu Ming" uniqKey="Wang Y" first="Yu-Ming" last="Wang">Yu-Ming Wang</name><name sortKey="Yeh, Yi Chien" sort="Yeh, Yi Chien" uniqKey="Yeh Y" first="Yi-Chien" last="Yeh">Yi-Chien Yeh</name></noCountry><country name="Taïwan"><noRegion><name sortKey="Huang, Jau Ling" sort="Huang, Jau Ling" uniqKey="Huang J" first="Jau-Ling" last="Huang">Jau-Ling Huang</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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