Simultaneous detection and high-throughput identification of a panel of RNA viruses causing respiratory tract infections
Identifieur interne : 003B25 ( Main/Exploration ); précédent : 003B24; suivant : 003B26Simultaneous detection and high-throughput identification of a panel of RNA viruses causing respiratory tract infections
Auteurs : HAIJING LI [États-Unis] ; Melinda A. Mccormac [États-Unis] ; R. Wray Estes [États-Unis] ; Susan E. Sefers [États-Unis] ; Ryan K. Dare [États-Unis] ; James D. Chappell [États-Unis] ; Dean D. Erdman [États-Unis] ; Peter F. Wright [États-Unis] ; Yi-Wei Tang [États-Unis]Source :
- Journal of clinical microbiology : (Print) [ 0095-1137 ] ; 2007.
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- Pascal (Inist)
English descriptors
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Abstract
Clinical presentations for viral respiratory tract infections are often nonspecific, and a rapid, high-throughput laboratory technique that can detect a panel of common viral pathogens is clinically desirable. We evaluated two multiplex reverse transcription-PCR (RT-PCR) products coupled with microarray-based systems for simultaneous detection of common respiratory tract viral pathogens. The NGEN respiratory virus analyte-specific assay (Nanogen, San Diego, CA) detects influenza A virus (Flu-A) and Flu-B, parainfluenza virus 1 (PIV-1), PIV-2, and PIV-3, and respiratory syncytial virus (RSV), while the ResPlex II assay (Genaco Biomedical Products, Inc., Huntsville, AL) detects Flu-A, Flu-B, PIV-1, PIV-2, PIV-3, PIV-4, RSV, human metapneumovirus (hMPV), rhinoviruses (RhVs), enteroviruses (EnVs), and severe acute respiratory syndrome (SARS) coronavirus (CoV). A total of 360 frozen respiratory specimens collected for a full year were tested, and results were compared to those obtained with a combined reference standard of cell culture and monoplex real-time TaqMan RT-PCR assays. NGEN and ResPlex II gave comparable sensitivities for Flu-A (82.8 to 86.2%), Flu-B (90.0 to 100.0%), PIV-1 (87.5 to 93.8%), PIV-3 (66.7 to 72.2%), and RSV (63.3 to 73.3%); both assays achieved excellent specificities (99.1 to 100.0%) for these five common viruses. The ResPlex II assay detected hMPV in 13 (3.6%) specimens, with a sensitivity of 80.0% and specificity of 99.7%. The ResPlex II assay also differentiated RSV-A and RSV-B and gave positive results for RhV and EnV in 31 (8.6%) and 19 (5.3%) specimens, respectively. PIV-2, PIN-4, and SARS CoV were not detected in the specimens tested. The two systems can process 80 (NGEN) and 96 (ResPlex II) tests per run, with a hands-on time of approximately 60 min and test turnaround times of 6 h (ResPlex II) and 9 h (NGEN). Multiple-panel testing detected an additional unsuspected 9 (3.4%) PIV-1 and 10 (3.7%) PIV-3 infections. While test sensitivities for RSV and PIV-3 need improvement, both the NGEN and ResPlex II assays provide user-friendly and high-throughput tools for simultaneous detection and identification of a panel of common respiratory viral pathogens in a single test format. The multiplex approach enhances diagnosis through detection of respiratory viral etiologic agents in cases in which the presence of the agent was not suspected and a test was not ordered by the clinicians.
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Erdman</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Respiratory and Gastroenteritis Viruses Branch, Division of Viral Diseases, Centers for Disease Control and Prevention</s1><s2>Atlanta, Georgia 30333</s2><s3>USA</s3><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Atlanta, Georgia 30333</wicri:noRegion></affiliation></author><author><name sortKey="Wright, Peter F" sort="Wright, Peter F" uniqKey="Wright P" first="Peter F." last="Wright">Peter F. 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Mccormac</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Department of Pathology, Vanderbilt University School of Medicine</s1><s2>Nashville, Tennessee 37232</s2><s3>USA</s3><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Nashville, Tennessee 37232</wicri:noRegion></affiliation></author><author><name sortKey="Wray Estes, R" sort="Wray Estes, R" uniqKey="Wray Estes R" first="R." last="Wray Estes">R. Wray Estes</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Department of Pathology, Vanderbilt University School of Medicine</s1><s2>Nashville, Tennessee 37232</s2><s3>USA</s3><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Nashville, Tennessee 37232</wicri:noRegion></affiliation></author><author><name sortKey="Sefers, Susan E" sort="Sefers, Susan E" uniqKey="Sefers S" first="Susan E." last="Sefers">Susan E. Sefers</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Department of Pathology, Vanderbilt University School of Medicine</s1><s2>Nashville, Tennessee 37232</s2><s3>USA</s3><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Nashville, Tennessee 37232</wicri:noRegion></affiliation></author><author><name sortKey="Dare, Ryan K" sort="Dare, Ryan K" uniqKey="Dare R" first="Ryan K." last="Dare">Ryan K. Dare</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Respiratory and Gastroenteritis Viruses Branch, Division of Viral Diseases, Centers for Disease Control and Prevention</s1><s2>Atlanta, Georgia 30333</s2><s3>USA</s3><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Atlanta, Georgia 30333</wicri:noRegion></affiliation></author><author><name sortKey="Chappell, James D" sort="Chappell, James D" uniqKey="Chappell J" first="James D." last="Chappell">James D. Chappell</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Department of Pathology, Vanderbilt University School of Medicine</s1><s2>Nashville, Tennessee 37232</s2><s3>USA</s3><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Nashville, Tennessee 37232</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Department of Pediatrics, Vanderbilt University School of Medicine</s1><s2>Nashville, Tennessee 37232</s2><s3>USA</s3><sZ>6 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Nashville, Tennessee 37232</wicri:noRegion></affiliation></author><author><name sortKey="Erdman, Dean D" sort="Erdman, Dean D" uniqKey="Erdman D" first="Dean D." last="Erdman">Dean D. Erdman</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Respiratory and Gastroenteritis Viruses Branch, Division of Viral Diseases, Centers for Disease Control and Prevention</s1><s2>Atlanta, Georgia 30333</s2><s3>USA</s3><sZ>5 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Atlanta, Georgia 30333</wicri:noRegion></affiliation></author><author><name sortKey="Wright, Peter F" sort="Wright, Peter F" uniqKey="Wright P" first="Peter F." last="Wright">Peter F. Wright</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Department of Pathology, Vanderbilt University School of Medicine</s1><s2>Nashville, Tennessee 37232</s2><s3>USA</s3><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Nashville, Tennessee 37232</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Department of Pediatrics, Vanderbilt University School of Medicine</s1><s2>Nashville, Tennessee 37232</s2><s3>USA</s3><sZ>6 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Nashville, Tennessee 37232</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="05"><s1>Department of Microbiology and Immunology, Vanderbilt University School of Medicine</s1><s2>Nashville, Tennessee 37232</s2><s3>USA</s3><sZ>8 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Nashville, Tennessee 37232</wicri:noRegion></affiliation></author><author><name sortKey="Tang, Yi Wei" sort="Tang, Yi Wei" uniqKey="Tang Y" first="Yi-Wei" last="Tang">Yi-Wei Tang</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Medicine, Vanderbilt University School of Medicine</s1><s2>Nashville, Tennessee 37232</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Nashville, Tennessee 37232</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Department of Pathology, Vanderbilt University School of Medicine</s1><s2>Nashville, Tennessee 37232</s2><s3>USA</s3><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ><sZ>9 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Nashville, Tennessee 37232</wicri:noRegion></affiliation></author></analytic><series><title level="j" type="main">Journal of clinical microbiology : (Print)</title><title level="j" type="abbreviated">J. clin. microbiol. : (Print)</title><idno type="ISSN">0095-1137</idno><imprint><date when="2007">2007</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Journal of clinical microbiology : (Print)</title><title level="j" type="abbreviated">J. clin. microbiol. : (Print)</title><idno type="ISSN">0095-1137</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Detection</term><term>High throughput screening</term><term>Identification</term><term>Method</term><term>Microbiology</term><term>RNA virus</term><term>Respiratory tract</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Détection</term><term>Criblage haut débit</term><term>Identification</term><term>Voie respiratoire</term><term>Microbiologie</term><term>Virus à ARN</term><term>Méthode</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Clinical presentations for viral respiratory tract infections are often nonspecific, and a rapid, high-throughput laboratory technique that can detect a panel of common viral pathogens is clinically desirable. We evaluated two multiplex reverse transcription-PCR (RT-PCR) products coupled with microarray-based systems for simultaneous detection of common respiratory tract viral pathogens. The NGEN respiratory virus analyte-specific assay (Nanogen, San Diego, CA) detects influenza A virus (Flu-A) and Flu-B, parainfluenza virus 1 (PIV-1), PIV-2, and PIV-3, and respiratory syncytial virus (RSV), while the ResPlex II assay (Genaco Biomedical Products, Inc., Huntsville, AL) detects Flu-A, Flu-B, PIV-1, PIV-2, PIV-3, PIV-4, RSV, human metapneumovirus (hMPV), rhinoviruses (RhVs), enteroviruses (EnVs), and severe acute respiratory syndrome (SARS) coronavirus (CoV). A total of 360 frozen respiratory specimens collected for a full year were tested, and results were compared to those obtained with a combined reference standard of cell culture and monoplex real-time TaqMan RT-PCR assays. NGEN and ResPlex II gave comparable sensitivities for Flu-A (82.8 to 86.2%), Flu-B (90.0 to 100.0%), PIV-1 (87.5 to 93.8%), PIV-3 (66.7 to 72.2%), and RSV (63.3 to 73.3%); both assays achieved excellent specificities (99.1 to 100.0%) for these five common viruses. The ResPlex II assay detected hMPV in 13 (3.6%) specimens, with a sensitivity of 80.0% and specificity of 99.7%. The ResPlex II assay also differentiated RSV-A and RSV-B and gave positive results for RhV and EnV in 31 (8.6%) and 19 (5.3%) specimens, respectively. PIV-2, PIN-4, and SARS CoV were not detected in the specimens tested. The two systems can process 80 (NGEN) and 96 (ResPlex II) tests per run, with a hands-on time of approximately 60 min and test turnaround times of 6 h (ResPlex II) and 9 h (NGEN). Multiple-panel testing detected an additional unsuspected 9 (3.4%) PIV-1 and 10 (3.7%) PIV-3 infections. While test sensitivities for RSV and PIV-3 need improvement, both the NGEN and ResPlex II assays provide user-friendly and high-throughput tools for simultaneous detection and identification of a panel of common respiratory viral pathogens in a single test format. The multiplex approach enhances diagnosis through detection of respiratory viral etiologic agents in cases in which the presence of the agent was not suspected and a test was not ordered by the clinicians.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country></list><tree><country name="États-Unis"><noRegion><name sortKey="Haijing Li" sort="Haijing Li" uniqKey="Haijing Li" last="Haijing Li">HAIJING LI</name></noRegion><name sortKey="Chappell, James D" sort="Chappell, James D" uniqKey="Chappell J" first="James D." last="Chappell">James D. Chappell</name><name sortKey="Chappell, James D" sort="Chappell, James D" uniqKey="Chappell J" first="James D." last="Chappell">James D. Chappell</name><name sortKey="Dare, Ryan K" sort="Dare, Ryan K" uniqKey="Dare R" first="Ryan K." last="Dare">Ryan K. Dare</name><name sortKey="Erdman, Dean D" sort="Erdman, Dean D" uniqKey="Erdman D" first="Dean D." last="Erdman">Dean D. Erdman</name><name sortKey="Mccormac, Melinda A" sort="Mccormac, Melinda A" uniqKey="Mccormac M" first="Melinda A." last="Mccormac">Melinda A. Mccormac</name><name sortKey="Sefers, Susan E" sort="Sefers, Susan E" uniqKey="Sefers S" first="Susan E." last="Sefers">Susan E. Sefers</name><name sortKey="Tang, Yi Wei" sort="Tang, Yi Wei" uniqKey="Tang Y" first="Yi-Wei" last="Tang">Yi-Wei Tang</name><name sortKey="Tang, Yi Wei" sort="Tang, Yi Wei" uniqKey="Tang Y" first="Yi-Wei" last="Tang">Yi-Wei Tang</name><name sortKey="Wray Estes, R" sort="Wray Estes, R" uniqKey="Wray Estes R" first="R." last="Wray Estes">R. Wray Estes</name><name sortKey="Wright, Peter F" sort="Wright, Peter F" uniqKey="Wright P" first="Peter F." last="Wright">Peter F. Wright</name><name sortKey="Wright, Peter F" sort="Wright, Peter F" uniqKey="Wright P" first="Peter F." last="Wright">Peter F. Wright</name><name sortKey="Wright, Peter F" sort="Wright, Peter F" uniqKey="Wright P" first="Peter F." last="Wright">Peter F. Wright</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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