Sensitive detection of SARS coronavirus RNA by a novel asymmetric multiplex nested RT-PCR amplification coupled with oligonucleotide microarray hybridization.
Identifieur interne : 002547 ( PubMed/Corpus ); précédent : 002546; suivant : 002548Sensitive detection of SARS coronavirus RNA by a novel asymmetric multiplex nested RT-PCR amplification coupled with oligonucleotide microarray hybridization.
Auteurs : Zhi-Wei Zhang ; Yi-Ming Zhou ; Yan Zhang ; Yong Guo ; Sheng-Ce Tao ; Ze Li ; Qiong Zhang ; Jing ChengSource :
- Methods in molecular medicine [ 1543-1894 ] ; 2005.
English descriptors
- KwdEn :
- Oligonucleotide Array Sequence Analysis (instrumentation), Oligonucleotide Array Sequence Analysis (methods), RNA, Viral (analysis), Reverse Transcriptase Polymerase Chain Reaction (instrumentation), Reverse Transcriptase Polymerase Chain Reaction (methods), SARS Virus (genetics), Sensitivity and Specificity.
- MESH :
- chemical , analysis : RNA, Viral.
- genetics : SARS Virus.
- instrumentation : Oligonucleotide Array Sequence Analysis, Reverse Transcriptase Polymerase Chain Reaction.
- methods : Oligonucleotide Array Sequence Analysis, Reverse Transcriptase Polymerase Chain Reaction.
- Sensitivity and Specificity.
Abstract
We have developed a sensitive method for the detection of specific genes simultaneously. First, DNA was amplified by a novel asymmetric multiplex PCR with universal primer(s). Second, the 6-carboxytetramethylrhodamine (TAMRA)-labeled PCR products were hybridized specifically with oligonucleotide microarrays. Finally, matched duplexes were detected by using a laser-induced fluorescence scanner. The usefulness of this method was illustrated by analyzing severe acute respiratory syndrome (SARS) coronavirus RNA. The detection limit was 10(0) copies/microL. The results of the asymmetric multiplex nested reverse transcription-PCR were in agreement with the results of the microarray hybridization; no hybridization signal was lost as happened with applicons from symmetric amplifications. This reliable method can be used to the identification of other microorganisms, screening of genetic diseases, and other applications.
DOI: 10.1385/1-59259-923-0:59
PubMed: 16156097
Links to Exploration step
pubmed:16156097Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Sensitive detection of SARS coronavirus RNA by a novel asymmetric multiplex nested RT-PCR amplification coupled with oligonucleotide microarray hybridization.</title><author><name sortKey="Zhang, Zhi Wei" sort="Zhang, Zhi Wei" uniqKey="Zhang Z" first="Zhi-Wei" last="Zhang">Zhi-Wei Zhang</name><affiliation><nlm:affiliation>Department of Biiological Sciences and Biotechnology, Tsinghua University, Beijing, China.</nlm:affiliation></affiliation></author><author><name sortKey="Zhou, Yi Ming" sort="Zhou, Yi Ming" uniqKey="Zhou Y" first="Yi-Ming" last="Zhou">Yi-Ming Zhou</name></author><author><name sortKey="Zhang, Yan" sort="Zhang, Yan" uniqKey="Zhang Y" first="Yan" last="Zhang">Yan Zhang</name></author><author><name sortKey="Guo, Yong" sort="Guo, Yong" uniqKey="Guo Y" first="Yong" last="Guo">Yong Guo</name></author><author><name sortKey="Tao, Sheng Ce" sort="Tao, Sheng Ce" uniqKey="Tao S" first="Sheng-Ce" last="Tao">Sheng-Ce Tao</name></author><author><name sortKey="Li, Ze" sort="Li, Ze" uniqKey="Li Z" first="Ze" last="Li">Ze Li</name></author><author><name sortKey="Zhang, Qiong" sort="Zhang, Qiong" uniqKey="Zhang Q" first="Qiong" last="Zhang">Qiong Zhang</name></author><author><name sortKey="Cheng, Jing" sort="Cheng, Jing" uniqKey="Cheng J" first="Jing" last="Cheng">Jing Cheng</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2005">2005</date><idno type="RBID">pubmed:16156097</idno><idno type="pmid">16156097</idno><idno type="doi">10.1385/1-59259-923-0:59</idno><idno type="wicri:Area/PubMed/Corpus">002547</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002547</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Sensitive detection of SARS coronavirus RNA by a novel asymmetric multiplex nested RT-PCR amplification coupled with oligonucleotide microarray hybridization.</title><author><name sortKey="Zhang, Zhi Wei" sort="Zhang, Zhi Wei" uniqKey="Zhang Z" first="Zhi-Wei" last="Zhang">Zhi-Wei Zhang</name><affiliation><nlm:affiliation>Department of Biiological Sciences and Biotechnology, Tsinghua University, Beijing, China.</nlm:affiliation></affiliation></author><author><name sortKey="Zhou, Yi Ming" sort="Zhou, Yi Ming" uniqKey="Zhou Y" first="Yi-Ming" last="Zhou">Yi-Ming Zhou</name></author><author><name sortKey="Zhang, Yan" sort="Zhang, Yan" uniqKey="Zhang Y" first="Yan" last="Zhang">Yan Zhang</name></author><author><name sortKey="Guo, Yong" sort="Guo, Yong" uniqKey="Guo Y" first="Yong" last="Guo">Yong Guo</name></author><author><name sortKey="Tao, Sheng Ce" sort="Tao, Sheng Ce" uniqKey="Tao S" first="Sheng-Ce" last="Tao">Sheng-Ce Tao</name></author><author><name sortKey="Li, Ze" sort="Li, Ze" uniqKey="Li Z" first="Ze" last="Li">Ze Li</name></author><author><name sortKey="Zhang, Qiong" sort="Zhang, Qiong" uniqKey="Zhang Q" first="Qiong" last="Zhang">Qiong Zhang</name></author><author><name sortKey="Cheng, Jing" sort="Cheng, Jing" uniqKey="Cheng J" first="Jing" last="Cheng">Jing Cheng</name></author></analytic><series><title level="j">Methods in molecular medicine</title><idno type="ISSN">1543-1894</idno><imprint><date when="2005" type="published">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Oligonucleotide Array Sequence Analysis (instrumentation)</term><term>Oligonucleotide Array Sequence Analysis (methods)</term><term>RNA, Viral (analysis)</term><term>Reverse Transcriptase Polymerase Chain Reaction (instrumentation)</term><term>Reverse Transcriptase Polymerase Chain Reaction (methods)</term><term>SARS Virus (genetics)</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>RNA, Viral</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="instrumentation" xml:lang="en"><term>Oligonucleotide Array Sequence Analysis</term><term>Reverse Transcriptase Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Oligonucleotide Array Sequence Analysis</term><term>Reverse Transcriptase Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Sensitivity and Specificity</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">We have developed a sensitive method for the detection of specific genes simultaneously. First, DNA was amplified by a novel asymmetric multiplex PCR with universal primer(s). Second, the 6-carboxytetramethylrhodamine (TAMRA)-labeled PCR products were hybridized specifically with oligonucleotide microarrays. Finally, matched duplexes were detected by using a laser-induced fluorescence scanner. The usefulness of this method was illustrated by analyzing severe acute respiratory syndrome (SARS) coronavirus RNA. The detection limit was 10(0) copies/microL. The results of the asymmetric multiplex nested reverse transcription-PCR were in agreement with the results of the microarray hybridization; no hybridization signal was lost as happened with applicons from symmetric amplifications. This reliable method can be used to the identification of other microorganisms, screening of genetic diseases, and other applications.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">16156097</PMID><DateCompleted><Year>2005</Year><Month>10</Month><Day>06</Day></DateCompleted><DateRevised><Year>2020</Year><Month>04</Month><Day>07</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">1543-1894</ISSN><JournalIssue CitedMedium="Print"><Volume>114</Volume><PubDate><Year>2005</Year></PubDate></JournalIssue><Title>Methods in molecular medicine</Title><ISOAbbreviation>Methods Mol. Med.</ISOAbbreviation></Journal><ArticleTitle>Sensitive detection of SARS coronavirus RNA by a novel asymmetric multiplex nested RT-PCR amplification coupled with oligonucleotide microarray hybridization.</ArticleTitle><Pagination><MedlinePgn>59-78</MedlinePgn></Pagination><Abstract><AbstractText>We have developed a sensitive method for the detection of specific genes simultaneously. First, DNA was amplified by a novel asymmetric multiplex PCR with universal primer(s). Second, the 6-carboxytetramethylrhodamine (TAMRA)-labeled PCR products were hybridized specifically with oligonucleotide microarrays. Finally, matched duplexes were detected by using a laser-induced fluorescence scanner. The usefulness of this method was illustrated by analyzing severe acute respiratory syndrome (SARS) coronavirus RNA. The detection limit was 10(0) copies/microL. The results of the asymmetric multiplex nested reverse transcription-PCR were in agreement with the results of the microarray hybridization; no hybridization signal was lost as happened with applicons from symmetric amplifications. This reliable method can be used to the identification of other microorganisms, screening of genetic diseases, and other applications.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Zhi-wei</ForeName><Initials>ZW</Initials><AffiliationInfo><Affiliation>Department of Biiological Sciences and Biotechnology, Tsinghua University, Beijing, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Zhou</LastName><ForeName>Yi-ming</ForeName><Initials>YM</Initials></Author><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Yan</ForeName><Initials>Y</Initials></Author><Author ValidYN="Y"><LastName>Guo</LastName><ForeName>Yong</ForeName><Initials>Y</Initials></Author><Author ValidYN="Y"><LastName>Tao</LastName><ForeName>Sheng-ce</ForeName><Initials>SC</Initials></Author><Author ValidYN="Y"><LastName>Li</LastName><ForeName>Ze</ForeName><Initials>Z</Initials></Author><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Qiong</ForeName><Initials>Q</Initials></Author><Author ValidYN="Y"><LastName>Cheng</LastName><ForeName>Jing</ForeName><Initials>J</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>Methods Mol Med</MedlineTA><NlmUniqueID>101123138</NlmUniqueID><ISSNLinking>1543-1894</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D012367">RNA, Viral</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D020411" MajorTopicYN="Y">Oligonucleotide Array Sequence Analysis</DescriptorName><QualifierName UI="Q000295" 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