Development of a quantitative assay for SARS coronavirus and correlation of GAPDH mRNA with SARS coronavirus in clinical specimens
Identifieur interne : 000764 ( Istex/Corpus ); précédent : 000763; suivant : 000765Development of a quantitative assay for SARS coronavirus and correlation of GAPDH mRNA with SARS coronavirus in clinical specimens
Auteurs : S C C. Wong ; J K C. Chan ; K C Lee ; E S F. Lo ; D N C. TsangSource :
- Journal of Clinical Pathology [ 0021-9746 ] ; 2005-03.
English descriptors
- KwdEn :
- CoV, coronavirus, GAPDH, glyceraldehyde-3-phosphate dehydrogenase, Q-RT-PCR, quantitative reverse transcriptase polymerase chain reaction, RT-PCR, reverse transcriptase polymerase chain reaction, SARS, severe acute respiratory syndrome, WHO, World Health Organisation, cDNA, complementary deoxyribonucleic acid.
- Teeft :
- Actin, Actin mrna, Administrative region, Aliquot, Aspirate, Assay, Cdna, Clinical specimens, Coronavirus, Dehydrogenase, Detectable gapdh mrna, Gapdh, Gapdh mrna, Gapdh mrna concentration, Gapdh mrna concentrations, Gapdh mrna detection, Healthy controls, Healthy subjects, Higher amounts, Hong kong, Mrna, Nasopharyngeal, Nasopharyngeal swabs, Normal subjects, Polymerase, Positive controls, Primer, Queen elizabeth hospital, Rectal, Respiratory syndrome, Respiratory syndrome coronavirus, Sampling techniques, Sars, Sars coronavirus, Sars sars, Significant difference, Statistical analysis, Stool samples, Swab, Syndrome, Throat swabs, Transcriptase, Transcriptase polymerase chain reaction, Transcriptase polymerase chain reaction assay, Unrelated diseases, Urine, Urine specimens, World health organisation.
Abstract
Aims: To develop a quantitative reverse transcriptase polymerase chain reaction (Q-RT-PCR) for severe acute respiratory syndrome coronavirus (SARS-CoV) detection and explore the potential of using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA as an internal control to exclude false negative results. Methods: SARS-CoV and GAPDH mRNA were both measured in 26 specimens from 16 patients with SARS, 40 follow up specimens from the same batch of patients, and appropriate control subjects. The relation between SARS positivity and GAPDH mRNA concentration was investigated using the χ2 test. Increasing the sensitivity for SARS-CoV and GAPDH mRNA detection was investigated in follow up specimens in which SARS-CoV and GAPDH mRNA were not detected initially. Results: Varying amounts of SARS-CoV were found in the 26 SARS-CoV positive specimens and SARS-CoV was not detected in the 40 follow up specimens and controls. In addition, concentrations of GAPDH mRNA were significantly different between the patients with SARS, follow up specimens, and healthy controls (Kruskal-Wallis test, p<0.05). Moreover, GAPDH mRNA concentrations were highly correlated with SARS-CoV positivity (χ2 = 5.43; p<0.05). Finally, SARS-CoV and GAPDH mRNA were both detected in three follow up urine specimens that were initially negative when the amount of cDNA used was increased from 5 μl to 10 and 15 μl. Conclusions: This Q-RT-PCR assay can be used to detect SARS-CoV. Moreover, GAPDH mRNA may be useful to rule out false negative results in SARS-CoV detection, and the current extraction method for urine may not be sensitive enough to detect low titres of SARS-CoV.
Url:
DOI: 10.1136/jcp.2004.016592
Links to Exploration step
ISTEX:AE7CAE9B9EA1F3FDB3F1B19A0CD0D80DD8BA42AELe document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">Development of a quantitative assay for SARS coronavirus and correlation of GAPDH mRNA with SARS coronavirus in clinical specimens</title><author><name sortKey="Wong, S C C" sort="Wong, S C C" uniqKey="Wong S" first="S C C" last="Wong">S C C. Wong</name><affiliation><mods:affiliation>Department of Clinical Oncology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong Special Administrative Region, China</mods:affiliation></affiliation></author><author><name sortKey="Chan, J K C" sort="Chan, J K C" uniqKey="Chan J" first="J K C" last="Chan">J K C. Chan</name><affiliation><mods:affiliation>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</mods:affiliation></affiliation></author><author><name sortKey="Lee, K C" sort="Lee, K C" uniqKey="Lee K" first="K C" last="Lee">K C Lee</name><affiliation><mods:affiliation>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</mods:affiliation></affiliation></author><author><name sortKey="Lo, E S F" sort="Lo, E S F" uniqKey="Lo E" first="E S F" last="Lo">E S F. Lo</name><affiliation><mods:affiliation>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</mods:affiliation></affiliation></author><author><name sortKey="Tsang, D N C" sort="Tsang, D N C" uniqKey="Tsang D" first="D N C" last="Tsang">D N C. Tsang</name><affiliation><mods:affiliation>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</mods:affiliation></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:AE7CAE9B9EA1F3FDB3F1B19A0CD0D80DD8BA42AE</idno><date when="2005" year="2005">2005</date><idno type="doi">10.1136/jcp.2004.016592</idno><idno type="url">https://api.istex.fr/ark:/67375/NVC-CNFP4K1H-1/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">000764</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">000764</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main" xml:lang="en">Development of a quantitative assay for SARS coronavirus and correlation of GAPDH mRNA with SARS coronavirus in clinical specimens</title><author><name sortKey="Wong, S C C" sort="Wong, S C C" uniqKey="Wong S" first="S C C" last="Wong">S C C. Wong</name><affiliation><mods:affiliation>Department of Clinical Oncology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong Special Administrative Region, China</mods:affiliation></affiliation></author><author><name sortKey="Chan, J K C" sort="Chan, J K C" uniqKey="Chan J" first="J K C" last="Chan">J K C. Chan</name><affiliation><mods:affiliation>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</mods:affiliation></affiliation></author><author><name sortKey="Lee, K C" sort="Lee, K C" uniqKey="Lee K" first="K C" last="Lee">K C Lee</name><affiliation><mods:affiliation>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</mods:affiliation></affiliation></author><author><name sortKey="Lo, E S F" sort="Lo, E S F" uniqKey="Lo E" first="E S F" last="Lo">E S F. Lo</name><affiliation><mods:affiliation>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</mods:affiliation></affiliation></author><author><name sortKey="Tsang, D N C" sort="Tsang, D N C" uniqKey="Tsang D" first="D N C" last="Tsang">D N C. Tsang</name><affiliation><mods:affiliation>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Journal of Clinical Pathology</title><title level="j" type="abbrev">J Clin Pathol</title><idno type="ISSN">0021-9746</idno><idno type="eISSN">1472-4146</idno><imprint><publisher>BMJ Publishing Group Ltd and Association of Clinical Pathologists</publisher><date type="published" when="2005-03">2005-03</date><biblScope unit="volume">58</biblScope><biblScope unit="issue">3</biblScope><biblScope unit="page" from="276">276</biblScope></imprint><idno type="ISSN">0021-9746</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0021-9746</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>CoV, coronavirus</term><term>GAPDH, glyceraldehyde-3-phosphate dehydrogenase</term><term>Q-RT-PCR, quantitative reverse transcriptase polymerase chain reaction</term><term>RT-PCR, reverse transcriptase polymerase chain reaction</term><term>SARS, severe acute respiratory syndrome</term><term>WHO, World Health Organisation</term><term>cDNA, complementary deoxyribonucleic acid</term></keywords><keywords scheme="Teeft" xml:lang="en"><term>Actin</term><term>Actin mrna</term><term>Administrative region</term><term>Aliquot</term><term>Aspirate</term><term>Assay</term><term>Cdna</term><term>Clinical specimens</term><term>Coronavirus</term><term>Dehydrogenase</term><term>Detectable gapdh mrna</term><term>Gapdh</term><term>Gapdh mrna</term><term>Gapdh mrna concentration</term><term>Gapdh mrna concentrations</term><term>Gapdh mrna detection</term><term>Healthy controls</term><term>Healthy subjects</term><term>Higher amounts</term><term>Hong kong</term><term>Mrna</term><term>Nasopharyngeal</term><term>Nasopharyngeal swabs</term><term>Normal subjects</term><term>Polymerase</term><term>Positive controls</term><term>Primer</term><term>Queen elizabeth hospital</term><term>Rectal</term><term>Respiratory syndrome</term><term>Respiratory syndrome coronavirus</term><term>Sampling techniques</term><term>Sars</term><term>Sars coronavirus</term><term>Sars sars</term><term>Significant difference</term><term>Statistical analysis</term><term>Stool samples</term><term>Swab</term><term>Syndrome</term><term>Throat swabs</term><term>Transcriptase</term><term>Transcriptase polymerase chain reaction</term><term>Transcriptase polymerase chain reaction assay</term><term>Unrelated diseases</term><term>Urine</term><term>Urine specimens</term><term>World health organisation</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Aims: To develop a quantitative reverse transcriptase polymerase chain reaction (Q-RT-PCR) for severe acute respiratory syndrome coronavirus (SARS-CoV) detection and explore the potential of using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA as an internal control to exclude false negative results. Methods: SARS-CoV and GAPDH mRNA were both measured in 26 specimens from 16 patients with SARS, 40 follow up specimens from the same batch of patients, and appropriate control subjects. The relation between SARS positivity and GAPDH mRNA concentration was investigated using the χ2 test. Increasing the sensitivity for SARS-CoV and GAPDH mRNA detection was investigated in follow up specimens in which SARS-CoV and GAPDH mRNA were not detected initially. Results: Varying amounts of SARS-CoV were found in the 26 SARS-CoV positive specimens and SARS-CoV was not detected in the 40 follow up specimens and controls. In addition, concentrations of GAPDH mRNA were significantly different between the patients with SARS, follow up specimens, and healthy controls (Kruskal-Wallis test, p<0.05). Moreover, GAPDH mRNA concentrations were highly correlated with SARS-CoV positivity (χ2 = 5.43; p<0.05). Finally, SARS-CoV and GAPDH mRNA were both detected in three follow up urine specimens that were initially negative when the amount of cDNA used was increased from 5 μl to 10 and 15 μl. Conclusions: This Q-RT-PCR assay can be used to detect SARS-CoV. Moreover, GAPDH mRNA may be useful to rule out false negative results in SARS-CoV detection, and the current extraction method for urine may not be sensitive enough to detect low titres of SARS-CoV.</div></front></TEI><istex><corpusName>bmj</corpusName><keywords><teeft><json:string>mrna</json:string><json:string>gapdh</json:string><json:string>sars</json:string><json:string>gapdh mrna</json:string><json:string>coronavirus</json:string><json:string>cdna</json:string><json:string>nasopharyngeal</json:string><json:string>actin</json:string><json:string>primer</json:string><json:string>swab</json:string><json:string>assay</json:string><json:string>dehydrogenase</json:string><json:string>respiratory syndrome</json:string><json:string>actin mrna</json:string><json:string>aspirate</json:string><json:string>hong kong</json:string><json:string>transcriptase</json:string><json:string>polymerase</json:string><json:string>gapdh mrna concentrations</json:string><json:string>rectal</json:string><json:string>aliquot</json:string><json:string>gapdh mrna detection</json:string><json:string>gapdh mrna concentration</json:string><json:string>urine</json:string><json:string>unrelated diseases</json:string><json:string>urine specimens</json:string><json:string>nasopharyngeal swabs</json:string><json:string>world health organisation</json:string><json:string>throat swabs</json:string><json:string>transcriptase polymerase chain reaction</json:string><json:string>sars coronavirus</json:string><json:string>clinical specimens</json:string><json:string>administrative region</json:string><json:string>higher amounts</json:string><json:string>queen elizabeth hospital</json:string><json:string>syndrome</json:string><json:string>sars sars</json:string><json:string>positive controls</json:string><json:string>statistical analysis</json:string><json:string>significant difference</json:string><json:string>sampling techniques</json:string><json:string>stool samples</json:string><json:string>respiratory syndrome coronavirus</json:string><json:string>normal subjects</json:string><json:string>transcriptase polymerase chain reaction assay</json:string><json:string>healthy controls</json:string><json:string>detectable gapdh mrna</json:string><json:string>healthy subjects</json:string></teeft></keywords><author><json:item><name>S C C Wong</name><affiliations><json:string>Department of Clinical Oncology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong Special Administrative Region, China</json:string></affiliations></json:item><json:item><name>J K C Chan</name><affiliations><json:string>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</json:string></affiliations></json:item><json:item><name>K C Lee</name><affiliations><json:string>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</json:string></affiliations></json:item><json:item><name>E S F Lo</name><affiliations><json:string>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</json:string></affiliations></json:item><json:item><name>D N C Tsang</name><affiliations><json:string>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>cDNA, complementary deoxyribonucleic acid</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>CoV, coronavirus</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>GAPDH, glyceraldehyde-3-phosphate dehydrogenase</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Q-RT-PCR, quantitative reverse transcriptase polymerase chain reaction</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>RT-PCR, reverse transcriptase polymerase chain reaction</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>SARS, severe acute respiratory syndrome</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>WHO, World Health Organisation</value></json:item></subject><arkIstex>ark:/67375/NVC-CNFP4K1H-1</arkIstex><language><json:string>eng</json:string></language><originalGenre><json:string>research-article</json:string></originalGenre><abstract>Aims: To develop a quantitative reverse transcriptase polymerase chain reaction (Q-RT-PCR) for severe acute respiratory syndrome coronavirus (SARS-CoV) detection and explore the potential of using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA as an internal control to exclude false negative results. Methods: SARS-CoV and GAPDH mRNA were both measured in 26 specimens from 16 patients with SARS, 40 follow up specimens from the same batch of patients, and appropriate control subjects. The relation between SARS positivity and GAPDH mRNA concentration was investigated using the χ2 test. Increasing the sensitivity for SARS-CoV and GAPDH mRNA detection was investigated in follow up specimens in which SARS-CoV and GAPDH mRNA were not detected initially. Results: Varying amounts of SARS-CoV were found in the 26 SARS-CoV positive specimens and SARS-CoV was not detected in the 40 follow up specimens and controls. In addition, concentrations of GAPDH mRNA were significantly different between the patients with SARS, follow up specimens, and healthy controls (Kruskal-Wallis test, p>0.05). Moreover, GAPDH mRNA concentrations were highly correlated with SARS-CoV positivity (χ2 = 5.43; p>0.05). Finally, SARS-CoV and GAPDH mRNA were both detected in three follow up urine specimens that were initially negative when the amount of cDNA used was increased from 5 μl to 10 and 15 μl. Conclusions: This Q-RT-PCR assay can be used to detect SARS-CoV. Moreover, GAPDH mRNA may be useful to rule out false negative results in SARS-CoV detection, and the current extraction method for urine may not be sensitive enough to detect low titres of SARS-CoV.</abstract><qualityIndicators><score>8.452</score><pdfWordCount>3500</pdfWordCount><pdfCharCount>23224</pdfCharCount><pdfVersion>1.3</pdfVersion><pdfPageCount>5</pdfPageCount><pdfPageSize>612 x 792 pts (letter)</pdfPageSize><refBibsNative>true</refBibsNative><abstractWordCount>246</abstractWordCount><abstractCharCount>1654</abstractCharCount><keywordCount>7</keywordCount></qualityIndicators><title>Development of a quantitative assay for SARS coronavirus and correlation of GAPDH mRNA with SARS coronavirus in clinical specimens</title><pmid><json:string>15735160</json:string></pmid><genre><json:string>research-article</json:string></genre><host><title>Journal of Clinical Pathology</title><language><json:string>unknown</json:string></language><issn><json:string>0021-9746</json:string></issn><eissn><json:string>1472-4146</json:string></eissn><volume>58</volume><issue>3</issue><pages><first>276</first></pages><genre><json:string>journal</json:string></genre></host><namedEntities><unitex><date><json:string>2005-02-25</json:string></date><geogName></geogName><orgName><json:string>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</json:string><json:string>WHO</json:string><json:string>Life Technologies</json:string><json:string>World Health Organisation Quantitative SARS</json:string><json:string>Queen Elizabeth Hospital, Hong Kong, China</json:string><json:string>World Health Organisation</json:string><json:string>Department of Clinical Oncology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong Special Administrative Region, China J K C Chan</json:string><json:string>Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</json:string><json:string>the Pathology Department, Queen Elizabeth Hospital</json:string></orgName><orgName_funder><json:string>the Pathology Department, Queen Elizabeth Hospital</json:string></orgName_funder><orgName_provider></orgName_provider><persName><json:string>Cesar Wong</json:string><json:string>Patient</json:string></persName><placeName><json:string>Foster City</json:string><json:string>San Diego</json:string><json:string>Germany</json:string><json:string>Hong Kong</json:string><json:string>China</json:string></placeName><ref_url></ref_url><ref_bibl><json:string>April and June 2003</json:string><json:string>GraphPad Software Inc, 1999</json:string></ref_bibl><bibl></bibl></unitex></namedEntities><ark><json:string>ark:/67375/NVC-CNFP4K1H-1</json:string></ark><categories><wos><json:string>1 - science</json:string><json:string>2 - pathology</json:string></wos><scienceMetrix></scienceMetrix><scopus><json:string>1 - Health Sciences</json:string><json:string>2 - Medicine</json:string><json:string>3 - General Medicine</json:string><json:string>1 - Health Sciences</json:string><json:string>2 - Medicine</json:string><json:string>3 - Pathology and Forensic Medicine</json:string></scopus><inist><json:string>1 - sciences appliquees, technologies et medecines</json:string><json:string>2 - sciences biologiques et medicales</json:string><json:string>3 - sciences medicales</json:string></inist></categories><publicationDate>2005</publicationDate><copyrightDate>2005</copyrightDate><doi><json:string>10.1136/jcp.2004.016592</json:string></doi><id>AE7CAE9B9EA1F3FDB3F1B19A0CD0D80DD8BA42AE</id><score>1</score><fulltext><json:item><extension>pdf</extension><original>true</original><mimetype>application/pdf</mimetype><uri>https://api.istex.fr/ark:/67375/NVC-CNFP4K1H-1/fulltext.pdf</uri></json:item><json:item><extension>zip</extension><original>false</original><mimetype>application/zip</mimetype><uri>https://api.istex.fr/ark:/67375/NVC-CNFP4K1H-1/bundle.zip</uri></json:item><istex:fulltextTEI uri="https://api.istex.fr/ark:/67375/NVC-CNFP4K1H-1/fulltext.tei"><teiHeader><fileDesc><titleStmt><title level="a" type="main" xml:lang="en">Development of a quantitative assay for SARS coronavirus and correlation of GAPDH mRNA with SARS coronavirus in clinical specimens</title></titleStmt><publicationStmt><authority>ISTEX</authority><publisher scheme="https://publisher-list.data.istex.fr">BMJ Publishing Group Ltd and Association of Clinical Pathologists</publisher><availability><licence><p>Copyright 2005 Journal of Clinical Pathology</p></licence><p scheme="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-7M42M2QJ-2">bmj</p></availability><date>2005-02-25</date></publicationStmt><notesStmt><note type="research-article" scheme="https://content-type.data.istex.fr/ark:/67375/XTP-1JC4F85T-7">research-article</note><note type="journal" scheme="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</note><note>Correspondence to:
Dr S C C Wong
Department of Clinical Oncology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong Special Administrative Region, China; cesar@clo.cuhk.edu.hk</note></notesStmt><sourceDesc><biblStruct type="inbook"><analytic><title level="a" type="main" xml:lang="en">Development of a quantitative assay for SARS coronavirus and correlation of GAPDH mRNA with SARS coronavirus in clinical specimens</title><author xml:id="author-0000"><persName><forename type="first">S C C</forename><surname>Wong</surname></persName><affiliation>Department of Clinical Oncology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong Special Administrative Region, China</affiliation></author><author xml:id="author-0001"><persName><forename type="first">J K C</forename><surname>Chan</surname></persName><affiliation>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</affiliation></author><author xml:id="author-0002"><persName><forename type="first">K C</forename><surname>Lee</surname></persName><affiliation>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</affiliation></author><author xml:id="author-0003"><persName><forename type="first">E S F</forename><surname>Lo</surname></persName><affiliation>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</affiliation></author><author xml:id="author-0004"><persName><forename type="first">D N C</forename><surname>Tsang</surname></persName><affiliation>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</affiliation></author><idno type="istex">AE7CAE9B9EA1F3FDB3F1B19A0CD0D80DD8BA42AE</idno><idno type="ark">ark:/67375/NVC-CNFP4K1H-1</idno><idno type="DOI">10.1136/jcp.2004.016592</idno><idno type="href">jclinpath-58-276.pdf</idno><idno type="PMID">15735160</idno><idno type="local">0580276</idno></analytic><monogr><title level="j">Journal of Clinical Pathology</title><title level="j" type="abbrev">J Clin Pathol</title><idno type="pISSN">0021-9746</idno><idno type="eISSN">1472-4146</idno><idno type="PublisherID-hwp">jclinpath</idno><idno type="PublisherID-nlm-ta">J Clin Pathol</idno><imprint><publisher>BMJ Publishing Group Ltd and Association of Clinical Pathologists</publisher><date type="published" when="2005-03"></date><biblScope unit="volume">58</biblScope><biblScope unit="issue">3</biblScope><biblScope unit="page" from="276">276</biblScope></imprint></monogr></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>2005-02-25</date></creation><langUsage><language ident="en">en</language></langUsage><abstract xml:lang="en"><p>Aims: To develop a quantitative reverse transcriptase polymerase chain reaction (Q-RT-PCR) for severe acute respiratory syndrome coronavirus (SARS-CoV) detection and explore the potential of using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA as an internal control to exclude false negative results. Methods: SARS-CoV and GAPDH mRNA were both measured in 26 specimens from 16 patients with SARS, 40 follow up specimens from the same batch of patients, and appropriate control subjects. The relation between SARS positivity and GAPDH mRNA concentration was investigated using the χ2 test. Increasing the sensitivity for SARS-CoV and GAPDH mRNA detection was investigated in follow up specimens in which SARS-CoV and GAPDH mRNA were not detected initially. Results: Varying amounts of SARS-CoV were found in the 26 SARS-CoV positive specimens and SARS-CoV was not detected in the 40 follow up specimens and controls. In addition, concentrations of GAPDH mRNA were significantly different between the patients with SARS, follow up specimens, and healthy controls (Kruskal-Wallis test, p<0.05). Moreover, GAPDH mRNA concentrations were highly correlated with SARS-CoV positivity (χ2 = 5.43; p<0.05). Finally, SARS-CoV and GAPDH mRNA were both detected in three follow up urine specimens that were initially negative when the amount of cDNA used was increased from 5 μl to 10 and 15 μl. Conclusions: This Q-RT-PCR assay can be used to detect SARS-CoV. Moreover, GAPDH mRNA may be useful to rule out false negative results in SARS-CoV detection, and the current extraction method for urine may not be sensitive enough to detect low titres of SARS-CoV.</p></abstract><textClass xml:lang="en"><keywords scheme="keyword"><list><head>ABR</head><item><term>cDNA, complementary deoxyribonucleic acid</term></item><item><term>CoV, coronavirus</term></item><item><term>GAPDH, glyceraldehyde-3-phosphate dehydrogenase</term></item><item><term>Q-RT-PCR, quantitative reverse transcriptase polymerase chain reaction</term></item><item><term>RT-PCR, reverse transcriptase polymerase chain reaction</term></item><item><term>SARS, severe acute respiratory syndrome</term></item><item><term>WHO, World Health Organisation</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="2005-02-25">Created</change><change when="2005-03">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><extension>txt</extension><original>false</original><mimetype>text/plain</mimetype><uri>https://api.istex.fr/ark:/67375/NVC-CNFP4K1H-1/fulltext.txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="corpus bmj" wicri:toSee="no header"><istex:xmlDeclaration>version="1.0" encoding="UTF-8" standalone="no"</istex:xmlDeclaration><istex:docType PUBLIC="-//NLM//DTD Journal Archiving and Interchange DTD v2.3 20070202//EN" URI="archivearticle.dtd" name="istex:docType"></istex:docType><istex:document><article xml:lang="en" article-type="research-article"><front><journal-meta><journal-id journal-id-type="hwp">jclinpath</journal-id><journal-id journal-id-type="nlm-ta">J Clin Pathol</journal-id><journal-title>Journal of Clinical Pathology</journal-title><abbrev-journal-title abbrev-type="publisher">J Clin Pathol</abbrev-journal-title><issn pub-type="ppub">0021-9746</issn><issn pub-type="epub">1472-4146</issn><publisher><publisher-name>BMJ Publishing Group Ltd and Association of Clinical Pathologists</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="other">0580276</article-id><article-id pub-id-type="doi">10.1136/jcp.2004.016592</article-id><article-id pub-id-type="other">jclinpath;58/3/276</article-id><article-id pub-id-type="pmid">15735160</article-id><article-id pub-id-type="other">276</article-id><article-id pub-id-type="other">jcp.2004.016592</article-id><article-categories><subj-group subj-group-type="heading"><subject content-type="original">Original articles</subject></subj-group></article-categories><title-group><article-title>Development of a quantitative assay for SARS coronavirus and correlation of GAPDH mRNA with SARS coronavirus in clinical specimens</article-title></title-group><contrib-group><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Wong</surname><given-names>S C C</given-names></name><xref rid="AFF1">1</xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Chan</surname><given-names>J K C</given-names></name><xref rid="AFF2">2</xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Lee</surname><given-names>K C</given-names></name><xref rid="AFF2">2</xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Lo</surname><given-names>E S F</given-names></name><xref rid="AFF2">2</xref></contrib><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Tsang</surname><given-names>D N C</given-names></name><xref rid="AFF2">2</xref></contrib><aff id="AFF1"><label>1</label>Department of Clinical Oncology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong Special Administrative Region, China</aff><aff id="AFF2"><label>2</label>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</aff></contrib-group><author-notes><corresp>Correspondence to:
Dr S C C Wong
Department of Clinical Oncology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong Special Administrative Region, China; <ext-link xlink:href="cesarclo.cuhk.edu.hk" ext-link-type="email" xlink:type="simple">cesar@clo.cuhk.edu.hk</ext-link></corresp></author-notes><pub-date pub-type="ppub"><month>3</month><year>2005</year></pub-date><pub-date pub-type="epub"><day>25</day><month>2</month><year>2005</year></pub-date><volume>58</volume><volume-id pub-id-type="other">58</volume-id><volume-id pub-id-type="other">58</volume-id><issue>3</issue><issue-id pub-id-type="other">jclinpath;58/3</issue-id><issue-id pub-id-type="other">3</issue-id><issue-id pub-id-type="other">58/3</issue-id><fpage>276</fpage><history><date date-type="accepted"><day>10</day><month>09</month><year>2004</year></date></history><permissions><copyright-statement>Copyright 2005 Journal of Clinical Pathology</copyright-statement><copyright-year>2005</copyright-year></permissions><self-uri content-type="pdf" xlink:role="full-text" xlink:href="jclinpath-58-276.pdf"></self-uri><abstract xml:lang="en"><p><bold>Aims:</bold> To develop a quantitative reverse transcriptase polymerase chain reaction (Q-RT-PCR) for severe acute respiratory syndrome coronavirus (SARS-CoV) detection and explore the potential of using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA as an internal control to exclude false negative results.</p><p><bold>Methods:</bold> SARS-CoV and GAPDH mRNA were both measured in 26 specimens from 16 patients with SARS, 40 follow up specimens from the same batch of patients, and appropriate control subjects. The relation between SARS positivity and GAPDH mRNA concentration was investigated using the χ<sup>2</sup> test. Increasing the sensitivity for SARS-CoV and GAPDH mRNA detection was investigated in follow up specimens in which SARS-CoV and GAPDH mRNA were not detected initially.</p><p><bold>Results:</bold> Varying amounts of SARS-CoV were found in the 26 SARS-CoV positive specimens and SARS-CoV was not detected in the 40 follow up specimens and controls. In addition, concentrations of GAPDH mRNA were significantly different between the patients with SARS, follow up specimens, and healthy controls (Kruskal-Wallis test, p<0.05). Moreover, GAPDH mRNA concentrations were highly correlated with SARS-CoV positivity (χ<sup>2</sup> = 5.43; p<0.05). Finally, SARS-CoV and GAPDH mRNA were both detected in three follow up urine specimens that were initially negative when the amount of cDNA used was increased from 5 μl to 10 and 15 μl.</p><p><bold>Conclusions:</bold> This Q-RT-PCR assay can be used to detect SARS-CoV. Moreover, GAPDH mRNA may be useful to rule out false negative results in SARS-CoV detection, and the current extraction method for urine may not be sensitive enough to detect low titres of SARS-CoV.</p></abstract><kwd-group kwd-group-type="ABR" xml:lang="en"><kwd>cDNA, complementary deoxyribonucleic acid</kwd><kwd>CoV, coronavirus</kwd><kwd>GAPDH, glyceraldehyde-3-phosphate dehydrogenase</kwd><kwd>Q-RT-PCR, quantitative reverse transcriptase polymerase chain reaction</kwd><kwd>RT-PCR, reverse transcriptase polymerase chain reaction</kwd><kwd>SARS, severe acute respiratory syndrome</kwd><kwd>WHO, World Health Organisation</kwd></kwd-group></article-meta></front></article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>Development of a quantitative assay for SARS coronavirus and correlation of GAPDH mRNA with SARS coronavirus in clinical specimens</title></titleInfo><titleInfo type="alternative" lang="en" contentType="CDATA"><title>Development of a quantitative assay for SARS coronavirus and correlation of GAPDH mRNA with SARS coronavirus in clinical specimens</title></titleInfo><name type="personal"><namePart type="given">S C C</namePart><namePart type="family">Wong</namePart><affiliation>Department of Clinical Oncology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong Special Administrative Region, China</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">J K C</namePart><namePart type="family">Chan</namePart><affiliation>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">K C</namePart><namePart type="family">Lee</namePart><affiliation>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">E S F</namePart><namePart type="family">Lo</namePart><affiliation>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">D N C</namePart><namePart type="family">Tsang</namePart><affiliation>Department of Pathology, Queen Elizabeth Hospital, Hong Kong Special Administrative Region, China</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="research-article" authority="ISTEX" authorityURI="https://content-type.data.istex.fr" valueURI="https://content-type.data.istex.fr/ark:/67375/XTP-1JC4F85T-7">research-article</genre><originInfo><publisher>BMJ Publishing Group Ltd and Association of Clinical Pathologists</publisher><dateIssued encoding="w3cdtf">2005-03</dateIssued><dateCreated encoding="w3cdtf">2005-02-25</dateCreated><copyrightDate encoding="w3cdtf">2005</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><abstract lang="en">Aims: To develop a quantitative reverse transcriptase polymerase chain reaction (Q-RT-PCR) for severe acute respiratory syndrome coronavirus (SARS-CoV) detection and explore the potential of using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA as an internal control to exclude false negative results. Methods: SARS-CoV and GAPDH mRNA were both measured in 26 specimens from 16 patients with SARS, 40 follow up specimens from the same batch of patients, and appropriate control subjects. The relation between SARS positivity and GAPDH mRNA concentration was investigated using the χ2 test. Increasing the sensitivity for SARS-CoV and GAPDH mRNA detection was investigated in follow up specimens in which SARS-CoV and GAPDH mRNA were not detected initially. Results: Varying amounts of SARS-CoV were found in the 26 SARS-CoV positive specimens and SARS-CoV was not detected in the 40 follow up specimens and controls. In addition, concentrations of GAPDH mRNA were significantly different between the patients with SARS, follow up specimens, and healthy controls (Kruskal-Wallis test, p<0.05). Moreover, GAPDH mRNA concentrations were highly correlated with SARS-CoV positivity (χ2 = 5.43; p<0.05). Finally, SARS-CoV and GAPDH mRNA were both detected in three follow up urine specimens that were initially negative when the amount of cDNA used was increased from 5 μl to 10 and 15 μl. Conclusions: This Q-RT-PCR assay can be used to detect SARS-CoV. Moreover, GAPDH mRNA may be useful to rule out false negative results in SARS-CoV detection, and the current extraction method for urine may not be sensitive enough to detect low titres of SARS-CoV.</abstract><note type="author-notes">Correspondence to:
Dr S C C Wong
Department of Clinical Oncology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong Special Administrative Region, China; cesar@clo.cuhk.edu.hk</note><subject lang="en"><genre>ABR</genre><topic>cDNA, complementary deoxyribonucleic acid</topic><topic>CoV, coronavirus</topic><topic>GAPDH, glyceraldehyde-3-phosphate dehydrogenase</topic><topic>Q-RT-PCR, quantitative reverse transcriptase polymerase chain reaction</topic><topic>RT-PCR, reverse transcriptase polymerase chain reaction</topic><topic>SARS, severe acute respiratory syndrome</topic><topic>WHO, World Health Organisation</topic></subject><relatedItem type="host"><titleInfo><title>Journal of Clinical Pathology</title></titleInfo><titleInfo type="abbreviated"><title>J Clin Pathol</title></titleInfo><genre type="journal" authority="ISTEX" authorityURI="https://publication-type.data.istex.fr" valueURI="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</genre><identifier type="ISSN">0021-9746</identifier><identifier type="eISSN">1472-4146</identifier><identifier type="PublisherID-hwp">jclinpath</identifier><identifier type="PublisherID-nlm-ta">J Clin Pathol</identifier><part><date>2005</date><detail type="volume"><caption>vol.</caption><number>58</number></detail><detail type="issue"><caption>no.</caption><number>3</number></detail><extent unit="pages"><start>276</start></extent></part></relatedItem><identifier type="istex">AE7CAE9B9EA1F3FDB3F1B19A0CD0D80DD8BA42AE</identifier><identifier type="ark">ark:/67375/NVC-CNFP4K1H-1</identifier><identifier type="DOI">10.1136/jcp.2004.016592</identifier><identifier type="href">jclinpath-58-276.pdf</identifier><identifier type="PMID">15735160</identifier><identifier type="local">0580276</identifier><accessCondition type="use and reproduction" contentType="copyright">Copyright 2005 Journal of Clinical Pathology</accessCondition><recordInfo><recordContentSource authority="ISTEX" authorityURI="https://loaded-corpus.data.istex.fr" valueURI="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-7M42M2QJ-2">bmj</recordContentSource><recordOrigin>Copyright 2005 Journal of Clinical Pathology</recordOrigin></recordInfo></mods><json:item><extension>json</extension><original>false</original><mimetype>application/json</mimetype><uri>https://api.istex.fr/ark:/67375/NVC-CNFP4K1H-1/record.json</uri></json:item></metadata><annexes><json:item><extension>jpeg</extension><original>true</original><mimetype>image/jpeg</mimetype><uri>https://api.istex.fr/ark:/67375/NVC-CNFP4K1H-1/annexes.jpeg</uri></json:item></annexes><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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