Development of Taqman RT-nested PCR system for clinical SARS-CoV detection.
Identifieur interne : 002E32 ( PubMed/Corpus ); précédent : 002E31; suivant : 002E33Development of Taqman RT-nested PCR system for clinical SARS-CoV detection.
Auteurs : Qingfa Wu ; Zuyuan Xu ; Tian Wei ; Haipang Zeng ; Jingxiang Li ; Haixue Gang ; Min Sun ; Fangbo Jiang ; Xiang Wang ; Wei Dong ; Ling Yang ; Jian WangSource :
- Journal of virological methods [ 0166-0934 ] ; 2004.
English descriptors
- KwdEn :
- Adolescent, Adult, Aged, Base Sequence, Child, China, DNA Primers (genetics), DNA, Viral (genetics), Female, Genome, Viral, Humans, Male, Middle Aged, Reverse Transcriptase Polymerase Chain Reaction (methods), Reverse Transcriptase Polymerase Chain Reaction (statistics & numerical data), SARS Virus (genetics), SARS Virus (isolation & purification), Sensitivity and Specificity, Severe Acute Respiratory Syndrome (diagnosis), Severe Acute Respiratory Syndrome (virology), Virology (methods), Virology (statistics & numerical data).
- MESH :
- chemical , genetics : DNA Primers, DNA, Viral.
- geographic : China.
- diagnosis : Severe Acute Respiratory Syndrome.
- genetics : SARS Virus.
- isolation & purification : SARS Virus.
- methods : Reverse Transcriptase Polymerase Chain Reaction, Virology.
- statistics & numerical data : Reverse Transcriptase Polymerase Chain Reaction, Virology.
- virology : Severe Acute Respiratory Syndrome.
- Adolescent, Adult, Aged, Base Sequence, Child, Female, Genome, Viral, Humans, Male, Middle Aged, Sensitivity and Specificity.
Abstract
Severe acute respiratory syndrome (SARS) is an acute newly emerged infectious respiratory illness. The etiologic agent of SARS was named 'SARS-associated coronavirus' (SARS-CoV) that can be detected with reverse transcription-polymerase chain reaction (RT-PCR) assays. In this study, 12 sets of nested primers covering the SARS-CoV genome have been screened and showed sufficient sensitivity to detect SARS-CoV in RNA isolated from virus cultured in Vero 6 cells. To optimize further the reaction condition of those nested primers sets, seven sets of nested primers have been chosen to compare their reverse transcribed efficiency with specific and random primers, which is useful to combine RT with the first round of PCR into a one-step RT-PCR. Based on the sensitivity and simplicity of results, the no. 73 primer set was chosen as the candidate primer set for clinical diagnoses. To specify the amplicon to minimize false positive results, a Taqman RT-nested PCR system of no. 73 nested primer set was developed. Through investigations on a test panel of whole blood obtained from 30 SARS patients and 9 control persons, the specificity and sensitivity of the Taqman RT-nested PCR system was found to be 100 and 83%, respectively, which suggests that the method is a promising one to diagnose SARS in early stages.
DOI: 10.1016/j.jviromet.2004.02.011
PubMed: 15109816
Links to Exploration step
pubmed:15109816Le document en format XML
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uniqKey="Gang H" first="Haixue" last="Gang">Haixue Gang</name></author><author><name sortKey="Sun, Min" sort="Sun, Min" uniqKey="Sun M" first="Min" last="Sun">Min Sun</name></author><author><name sortKey="Jiang, Fangbo" sort="Jiang, Fangbo" uniqKey="Jiang F" first="Fangbo" last="Jiang">Fangbo Jiang</name></author><author><name sortKey="Wang, Xiang" sort="Wang, Xiang" uniqKey="Wang X" first="Xiang" last="Wang">Xiang Wang</name></author><author><name sortKey="Dong, Wei" sort="Dong, Wei" uniqKey="Dong W" first="Wei" last="Dong">Wei Dong</name></author><author><name sortKey="Yang, Ling" sort="Yang, Ling" uniqKey="Yang L" first="Ling" last="Yang">Ling Yang</name></author><author><name sortKey="Wang, Jian" sort="Wang, Jian" uniqKey="Wang J" first="Jian" last="Wang">Jian Wang</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2004">2004</date><idno type="RBID">pubmed:15109816</idno><idno type="pmid">15109816</idno><idno 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Zeng</name></author><author><name sortKey="Li, Jingxiang" sort="Li, Jingxiang" uniqKey="Li J" first="Jingxiang" last="Li">Jingxiang Li</name></author><author><name sortKey="Gang, Haixue" sort="Gang, Haixue" uniqKey="Gang H" first="Haixue" last="Gang">Haixue Gang</name></author><author><name sortKey="Sun, Min" sort="Sun, Min" uniqKey="Sun M" first="Min" last="Sun">Min Sun</name></author><author><name sortKey="Jiang, Fangbo" sort="Jiang, Fangbo" uniqKey="Jiang F" first="Fangbo" last="Jiang">Fangbo Jiang</name></author><author><name sortKey="Wang, Xiang" sort="Wang, Xiang" uniqKey="Wang X" first="Xiang" last="Wang">Xiang Wang</name></author><author><name sortKey="Dong, Wei" sort="Dong, Wei" uniqKey="Dong W" first="Wei" last="Dong">Wei Dong</name></author><author><name sortKey="Yang, Ling" sort="Yang, Ling" uniqKey="Yang L" first="Ling" last="Yang">Ling Yang</name></author><author><name sortKey="Wang, Jian" sort="Wang, Jian" uniqKey="Wang J" first="Jian" last="Wang">Jian Wang</name></author></analytic><series><title level="j">Journal of virological methods</title><idno type="ISSN">0166-0934</idno><imprint><date when="2004" type="published">2004</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Adolescent</term><term>Adult</term><term>Aged</term><term>Base Sequence</term><term>Child</term><term>China</term><term>DNA Primers (genetics)</term><term>DNA, Viral (genetics)</term><term>Female</term><term>Genome, Viral</term><term>Humans</term><term>Male</term><term>Middle Aged</term><term>Reverse Transcriptase Polymerase Chain Reaction (methods)</term><term>Reverse Transcriptase Polymerase Chain Reaction (statistics & numerical data)</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><term>Virology (methods)</term><term>Virology (statistics & numerical data)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA Primers</term><term>DNA, Viral</term></keywords><keywords scheme="MESH" type="geographic" xml:lang="en"><term>China</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Reverse Transcriptase Polymerase Chain Reaction</term><term>Virology</term></keywords><keywords scheme="MESH" qualifier="statistics & numerical data" xml:lang="en"><term>Reverse Transcriptase Polymerase Chain Reaction</term><term>Virology</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Adolescent</term><term>Adult</term><term>Aged</term><term>Base Sequence</term><term>Child</term><term>Female</term><term>Genome, Viral</term><term>Humans</term><term>Male</term><term>Middle Aged</term><term>Sensitivity and Specificity</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Severe acute respiratory syndrome (SARS) is an acute newly emerged infectious respiratory illness. The etiologic agent of SARS was named 'SARS-associated coronavirus' (SARS-CoV) that can be detected with reverse transcription-polymerase chain reaction (RT-PCR) assays. In this study, 12 sets of nested primers covering the SARS-CoV genome have been screened and showed sufficient sensitivity to detect SARS-CoV in RNA isolated from virus cultured in Vero 6 cells. To optimize further the reaction condition of those nested primers sets, seven sets of nested primers have been chosen to compare their reverse transcribed efficiency with specific and random primers, which is useful to combine RT with the first round of PCR into a one-step RT-PCR. Based on the sensitivity and simplicity of results, the no. 73 primer set was chosen as the candidate primer set for clinical diagnoses. To specify the amplicon to minimize false positive results, a Taqman RT-nested PCR system of no. 73 nested primer set was developed. Through investigations on a test panel of whole blood obtained from 30 SARS patients and 9 control persons, the specificity and sensitivity of the Taqman RT-nested PCR system was found to be 100 and 83%, respectively, which suggests that the method is a promising one to diagnose SARS in early stages.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">15109816</PMID><DateCompleted><Year>2004</Year><Month>07</Month><Day>09</Day></DateCompleted><DateRevised><Year>2020</Year><Month>04</Month><Day>09</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0166-0934</ISSN><JournalIssue CitedMedium="Print"><Volume>119</Volume><Issue>1</Issue><PubDate><Year>2004</Year><Month>Jul</Month></PubDate></JournalIssue><Title>Journal of virological methods</Title><ISOAbbreviation>J. Virol. Methods</ISOAbbreviation></Journal><ArticleTitle>Development of Taqman RT-nested PCR system for clinical SARS-CoV detection.</ArticleTitle><Pagination><MedlinePgn>17-23</MedlinePgn></Pagination><Abstract><AbstractText>Severe acute respiratory syndrome (SARS) is an acute newly emerged infectious respiratory illness. The etiologic agent of SARS was named 'SARS-associated coronavirus' (SARS-CoV) that can be detected with reverse transcription-polymerase chain reaction (RT-PCR) assays. In this study, 12 sets of nested primers covering the SARS-CoV genome have been screened and showed sufficient sensitivity to detect SARS-CoV in RNA isolated from virus cultured in Vero 6 cells. To optimize further the reaction condition of those nested primers sets, seven sets of nested primers have been chosen to compare their reverse transcribed efficiency with specific and random primers, which is useful to combine RT with the first round of PCR into a one-step RT-PCR. Based on the sensitivity and simplicity of results, the no. 73 primer set was chosen as the candidate primer set for clinical diagnoses. To specify the amplicon to minimize false positive results, a Taqman RT-nested PCR system of no. 73 nested primer set was developed. Through investigations on a test panel of whole blood obtained from 30 SARS patients and 9 control persons, the specificity and sensitivity of the Taqman RT-nested PCR system was found to be 100 and 83%, respectively, which suggests that the method is a promising one to diagnose SARS in early stages.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Wu</LastName><ForeName>Qingfa</ForeName><Initials>Q</Initials><AffiliationInfo><Affiliation>Beijing Genomics Institute, Chinese Academy of Sciences, Airport Industrial Zone, Bldg 6, Beijing 101300, PR China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Xu</LastName><ForeName>Zuyuan</ForeName><Initials>Z</Initials></Author><Author ValidYN="Y"><LastName>Wei</LastName><ForeName>Tian</ForeName><Initials>T</Initials></Author><Author ValidYN="Y"><LastName>Zeng</LastName><ForeName>Haipang</ForeName><Initials>H</Initials></Author><Author ValidYN="Y"><LastName>Li</LastName><ForeName>Jingxiang</ForeName><Initials>J</Initials></Author><Author ValidYN="Y"><LastName>Gang</LastName><ForeName>Haixue</ForeName><Initials>H</Initials></Author><Author ValidYN="Y"><LastName>Sun</LastName><ForeName>Min</ForeName><Initials>M</Initials></Author><Author ValidYN="Y"><LastName>Jiang</LastName><ForeName>Fangbo</ForeName><Initials>F</Initials></Author><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Xiang</ForeName><Initials>X</Initials></Author><Author ValidYN="Y"><LastName>Dong</LastName><ForeName>Wei</ForeName><Initials>W</Initials></Author><Author ValidYN="Y"><LastName>Yang</LastName><ForeName>Ling</ForeName><Initials>L</Initials></Author><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Jian</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>Netherlands</Country><MedlineTA>J Virol Methods</MedlineTA><NlmUniqueID>8005839</NlmUniqueID><ISSNLinking>0166-0934</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D017931">DNA Primers</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D004279">DNA, Viral</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000293" MajorTopicYN="N">Adolescent</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D000328" MajorTopicYN="N">Adult</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D000368" MajorTopicYN="N">Aged</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D001483" MajorTopicYN="N">Base Sequence</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002648" 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IdType="pubmed">12595377</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>N Engl J Med. 2003 May 15;348(20):1967-76</Citation><ArticleIdList><ArticleId IdType="pubmed">12690091</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Chin Sci Bull. 2003;48(10):941-948</Citation><ArticleIdList><ArticleId IdType="pubmed">32214698</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Science. 2003 May 30;300(5624):1394-9</Citation><ArticleIdList><ArticleId IdType="pubmed">12730500</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Genome Res. 1996 Oct;6(10):986-94</Citation><ArticleIdList><ArticleId IdType="pubmed">8908518</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Clin Chem. 2003 Jun;49(6 Pt 1):953-5</Citation><ArticleIdList><ArticleId IdType="pubmed">12765993</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Anal Biochem. 2001 Feb 15;289(2):281-8</Citation><ArticleIdList><ArticleId IdType="pubmed">11161323</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Science. 2003 May 30;300(5624):1399-404</Citation><ArticleIdList><ArticleId IdType="pubmed">12730501</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Lancet. 2003 Apr 19;361(9366):1319-25</Citation><ArticleIdList><ArticleId IdType="pubmed">12711465</ArticleId></ArticleIdList></Reference></ReferenceList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
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