Multiplex SYBR Green Real-Time PCR Assay for Detection of Respiratory Viruses
Identifieur interne : 000023 ( Pmc/Curation ); précédent : 000022; suivant : 000024Multiplex SYBR Green Real-Time PCR Assay for Detection of Respiratory Viruses
Auteurs : Mozhdeh Sultani ; Talat Mokhtari Azad ; Mohammadreza Eshragian ; Azadeh Shadab ; Maryam Naseri ; Owrang Eilami ; Jila YavarianSource :
- Jundishapur Journal of Microbiology [ 2008-3645 ] ; 2015.
Abstract
It is often difficult for a physician to distinguish between viral and bacterial causes of respiratory infections and this may result in overuse of antibiotics. In many cases of community-acquired respiratory infections, clinicians treat patients empirically. The development of molecular methods for direct detection of viruses has been progressed recently.
The objective of this study was recognizing the panel of respiratory RNA viruses by multiplex SYBR Green real-time polymerase chain reaction (PCR).
Randomized 172 influenza-negative respiratory specimens of all age groups of hospitalized patients were collected. After RNA extraction, cDNA was synthesized. Three SYBR Green multiplex real-time PCR assays were developed for simultaneous detection of 12 respiratory RNA viruses. Each set of multiplex methods detected four viruses, the first set: respiratory syncytial virus, human metapneumovirus, rhinovirus, enterovirus; the second set: parainfluenza viruses 1 - 4 (PIV1-4); the third set: coronaviruses NL63, 229E, severe acute respiratory syndrome (SARS), and OC43.
Application of the multiplex SYBR Green real-time PCR in clinical samples from 172 patients in a one-year study resulted in detection of 19 (11.04%) PIV3, 9 (5.23%) PIV4, and 1 (0.58%) coronavirus NL63. All the positive samples were detected during December to March (2011 - 2012).
Multiplex SYBR Green real-time PCR is a rapid and relatively inexpensive method for detection of respiratory viruses.
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DOI: 10.5812/jjm.19041v2
PubMed: 26468358
PubMed Central: 4601230
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Mozhdeh Sultani<affiliation><nlm:aff id="aff63013">Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation><affiliation><nlm:aff id="aff63013">Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation><affiliation><nlm:aff id="aff63013">Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation><affiliation><nlm:aff id="aff63013">Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation><affiliation><nlm:aff id="aff63013">Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation><affiliation><nlm:aff id="aff63014">Department of Infectious Disease, Yasuj University of Medical Sciences, Yasuj, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation><affiliation><nlm:aff id="aff63013">Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation>Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Multiplex SYBR Green Real-Time PCR Assay for Detection of Respiratory Viruses</title><author><name sortKey="Sultani, Mozhdeh" sort="Sultani, Mozhdeh" uniqKey="Sultani M" first="Mozhdeh" last="Sultani">Mozhdeh Sultani</name><affiliation><nlm:aff id="aff63013">Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Mokhtari Azad, Talat" sort="Mokhtari Azad, Talat" uniqKey="Mokhtari Azad T" first="Talat" last="Mokhtari Azad">Talat Mokhtari Azad</name><affiliation><nlm:aff id="aff63013">Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Eshragian, Mohammadreza" sort="Eshragian, Mohammadreza" uniqKey="Eshragian M" first="Mohammadreza" last="Eshragian">Mohammadreza Eshragian</name><affiliation><nlm:aff id="aff63013">Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Shadab, Azadeh" sort="Shadab, Azadeh" uniqKey="Shadab A" first="Azadeh" last="Shadab">Azadeh Shadab</name><affiliation><nlm:aff id="aff63013">Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Naseri, Maryam" sort="Naseri, Maryam" uniqKey="Naseri M" first="Maryam" last="Naseri">Maryam Naseri</name><affiliation><nlm:aff id="aff63013">Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Eilami, Owrang" sort="Eilami, Owrang" uniqKey="Eilami O" first="Owrang" last="Eilami">Owrang Eilami</name><affiliation><nlm:aff id="aff63014">Department of Infectious Disease, Yasuj University of Medical Sciences, Yasuj, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Yavarian, Jila" sort="Yavarian, Jila" uniqKey="Yavarian J" first="Jila" last="Yavarian">Jila Yavarian</name><affiliation><nlm:aff id="aff63013">Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">26468358</idno><idno type="pmc">4601230</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4601230</idno><idno type="RBID">PMC:4601230</idno><idno type="doi">10.5812/jjm.19041v2</idno><date when="2015">2015</date><idno type="wicri:Area/Pmc/Corpus">000023</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000023</idno><idno type="wicri:Area/Pmc/Curation">000023</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000023</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Multiplex SYBR Green Real-Time PCR Assay for Detection of Respiratory Viruses</title><author><name sortKey="Sultani, Mozhdeh" sort="Sultani, Mozhdeh" uniqKey="Sultani M" first="Mozhdeh" last="Sultani">Mozhdeh Sultani</name><affiliation><nlm:aff id="aff63013">Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Mokhtari Azad, Talat" sort="Mokhtari Azad, Talat" uniqKey="Mokhtari Azad T" first="Talat" last="Mokhtari Azad">Talat Mokhtari Azad</name><affiliation><nlm:aff id="aff63013">Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Eshragian, Mohammadreza" sort="Eshragian, Mohammadreza" uniqKey="Eshragian M" first="Mohammadreza" last="Eshragian">Mohammadreza Eshragian</name><affiliation><nlm:aff id="aff63013">Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Shadab, Azadeh" sort="Shadab, Azadeh" uniqKey="Shadab A" first="Azadeh" last="Shadab">Azadeh Shadab</name><affiliation><nlm:aff id="aff63013">Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Naseri, Maryam" sort="Naseri, Maryam" uniqKey="Naseri M" first="Maryam" last="Naseri">Maryam Naseri</name><affiliation><nlm:aff id="aff63013">Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Eilami, Owrang" sort="Eilami, Owrang" uniqKey="Eilami O" first="Owrang" last="Eilami">Owrang Eilami</name><affiliation><nlm:aff id="aff63014">Department of Infectious Disease, Yasuj University of Medical Sciences, Yasuj, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author><author><name sortKey="Yavarian, Jila" sort="Yavarian, Jila" uniqKey="Yavarian J" first="Jila" last="Yavarian">Jila Yavarian</name><affiliation><nlm:aff id="aff63013">Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran</nlm:aff><wicri:noCountry code="subfield">IR Iran</wicri:noCountry></affiliation></author></analytic><series><title level="j">Jundishapur Journal of Microbiology</title><idno type="ISSN">2008-3645</idno><idno type="eISSN">2008-4161</idno><imprint><date when="2015">2015</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><sec><title>Background:</title><p>It is often difficult for a physician to distinguish between viral and bacterial causes of respiratory infections and this may result in overuse of antibiotics. In many cases of community-acquired respiratory infections, clinicians treat patients empirically. The development of molecular methods for direct detection of viruses has been progressed recently.</p></sec><sec><title>Objectives:</title><p>The objective of this study was recognizing the panel of respiratory RNA viruses by multiplex SYBR Green real-time polymerase chain reaction (PCR).</p></sec><sec><title>Materials and Methods:</title><p>Randomized 172 influenza-negative respiratory specimens of all age groups of hospitalized patients were collected. After RNA extraction, cDNA was synthesized. Three SYBR Green multiplex real-time PCR assays were developed for simultaneous detection of 12 respiratory RNA viruses. Each set of multiplex methods detected four viruses, the first set: respiratory syncytial virus, human metapneumovirus, rhinovirus, enterovirus; the second set: parainfluenza viruses 1 - 4 (PIV1-4); the third set: coronaviruses NL63, 229E, severe acute respiratory syndrome (SARS), and OC43.</p></sec><sec><title>Results:</title><p>Application of the multiplex SYBR Green real-time PCR in clinical samples from 172 patients in a one-year study resulted in detection of 19 (11.04%) PIV3, 9 (5.23%) PIV4, and 1 (0.58%) coronavirus NL63. All the positive samples were detected during December to March (2011 - 2012).</p></sec><sec><title>Conclusions:</title><p>Multiplex SYBR Green real-time PCR is a rapid and relatively inexpensive method for detection of respiratory viruses.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Pyrc, K" uniqKey="Pyrc K">K Pyrc</name></author><author><name sortKey="Stozek, K" uniqKey="Stozek K">K Stozek</name></author><author><name sortKey="Wojcik, K" uniqKey="Wojcik K">K Wojcik</name></author><author><name sortKey="Gawron, K" uniqKey="Gawron K">K Gawron</name></author><author><name sortKey="Zeglen, S" uniqKey="Zeglen S">S Zeglen</name></author><author><name sortKey="Karolak, W" uniqKey="Karolak W">W Karolak</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Girard, Mp" uniqKey="Girard M">MP Girard</name></author><author><name sortKey="Cherian, T" uniqKey="Cherian T">T 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Jundishapur J Microbiol</journal-id><journal-id journal-id-type="iso-abbrev">Jundishapur J Microbiol</journal-id><journal-id journal-id-type="doi">10.5812/jjm</journal-id><journal-id journal-id-type="publisher-id">Kowsar</journal-id><journal-title-group><journal-title>Jundishapur Journal of Microbiology</journal-title></journal-title-group><issn pub-type="ppub">2008-3645</issn><issn pub-type="epub">2008-4161</issn><publisher><publisher-name>Kowsar</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">26468358</article-id><article-id pub-id-type="pmc">4601230</article-id><article-id pub-id-type="doi">10.5812/jjm.19041v2</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject></subj-group></article-categories><title-group><article-title>Multiplex SYBR Green Real-Time PCR Assay for Detection of Respiratory Viruses</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Sultani</surname><given-names>Mozhdeh</given-names></name><xref ref-type="aff" rid="aff63013">1</xref></contrib><contrib contrib-type="author"><name><surname>Mokhtari Azad</surname><given-names>Talat</given-names></name><xref ref-type="aff" rid="aff63013">1</xref></contrib><contrib contrib-type="author"><name><surname>Eshragian</surname><given-names>Mohammadreza</given-names></name><xref ref-type="aff" rid="aff63013">1</xref></contrib><contrib contrib-type="author"><name><surname>Shadab</surname><given-names>Azadeh</given-names></name><xref ref-type="aff" rid="aff63013">1</xref></contrib><contrib contrib-type="author"><name><surname>Naseri</surname><given-names>Maryam</given-names></name><xref ref-type="aff" rid="aff63013">1</xref></contrib><contrib contrib-type="author"><name><surname>Eilami</surname><given-names>Owrang</given-names></name><xref ref-type="aff" rid="aff63014">2</xref></contrib><contrib contrib-type="author"><name><surname>Yavarian</surname><given-names>Jila</given-names></name><xref ref-type="aff" rid="aff63013">1</xref><xref ref-type="corresp" rid="cor63015">*</xref></contrib></contrib-group><aff id="aff63013"><label>1</label>Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran</aff><aff id="aff63014"><label>2</label>Department of Infectious Disease, Yasuj University of Medical Sciences, Yasuj, IR Iran</aff><author-notes><corresp id="cor63015"><label>*</label>Corresponding author: Jila Yavarian, Virology Department, Tehran University of Medical Sciences, Tehran, IR Iran. Tel: +98-2188962343, Fax: +98-2188962343, E-mail: <email>yavarian@tums.ac.ir</email></corresp></author-notes><pub-date pub-type="epub"><day>01</day><month>8</month><year>2015</year></pub-date><pub-date pub-type="collection"><month>8</month><year>2015</year></pub-date><volume>8</volume><issue>8</issue><elocation-id>e19041</elocation-id><history><date date-type="received"><day>18</day><month>3</month><year>2014</year></date><date date-type="rev-recd"><day>30</day><month>6</month><year>2014</year></date><date date-type="accepted"><day>25</day><month>7</month><year>2014</year></date></history><permissions><copyright-statement>Copyright © 2015, Ahvaz Jundishapur University of Medical Sciences.</copyright-statement><copyright-year>2015</copyright-year><license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by-nc/4.0/"><license-p><pmc-comment>CREATIVE COMMONS</pmc-comment>This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (<ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by-nc/4.0/">http://creativecommons.org/licenses/by-nc/4.0/</ext-link>) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.</license-p></license></permissions><abstract><sec><title>Background:</title><p>It is often difficult for a physician to distinguish between viral and bacterial causes of respiratory infections and this may result in overuse of antibiotics. In many cases of community-acquired respiratory infections, clinicians treat patients empirically. The development of molecular methods for direct detection of viruses has been progressed recently.</p></sec><sec><title>Objectives:</title><p>The objective of this study was recognizing the panel of respiratory RNA viruses by multiplex SYBR Green real-time polymerase chain reaction (PCR).</p></sec><sec><title>Materials and Methods:</title><p>Randomized 172 influenza-negative respiratory specimens of all age groups of hospitalized patients were collected. After RNA extraction, cDNA was synthesized. Three SYBR Green multiplex real-time PCR assays were developed for simultaneous detection of 12 respiratory RNA viruses. Each set of multiplex methods detected four viruses, the first set: respiratory syncytial virus, human metapneumovirus, rhinovirus, enterovirus; the second set: parainfluenza viruses 1 - 4 (PIV1-4); the third set: coronaviruses NL63, 229E, severe acute respiratory syndrome (SARS), and OC43.</p></sec><sec><title>Results:</title><p>Application of the multiplex SYBR Green real-time PCR in clinical samples from 172 patients in a one-year study resulted in detection of 19 (11.04%) PIV3, 9 (5.23%) PIV4, and 1 (0.58%) coronavirus NL63. All the positive samples were detected during December to March (2011 - 2012).</p></sec><sec><title>Conclusions:</title><p>Multiplex SYBR Green real-time PCR is a rapid and relatively inexpensive method for detection of respiratory viruses.</p></sec></abstract><kwd-group kwd-group-type="author"><kwd>SYBR Green</kwd><kwd>Real-Time PCR</kwd><kwd>Respiratory Syncytial Viruses</kwd></kwd-group></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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