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Identification of Pseudallescheria and Scedosporium Species by Three Molecular Methods
The major clinically relevant species in Scedosporium (teleomorph Pseudallescheria) are Pseudallescheria boydii, Scedosporium aurantiacum, Scedosporium apiospermum, and Scedosporium prolificans, while Pseudallescheria minutispora, Petriellopsis desertorum, and Scedosporium dehoogii are exceptional a...
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Published in: | Journal of Clinical Microbiology 2011-03, Vol.49 (3), p.960-967 |
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creator | Lu, Qiaoyun Gerrits van den Ende, A.H.G Bakkers, J.M.J.E Sun, Jiufeng Lackner, M Najafzadeh, M.J Melchers, W.J.G Li, Ruoyu de Hoog, G.S |
description | The major clinically relevant species in Scedosporium (teleomorph Pseudallescheria) are Pseudallescheria boydii, Scedosporium aurantiacum, Scedosporium apiospermum, and Scedosporium prolificans, while Pseudallescheria minutispora, Petriellopsis desertorum, and Scedosporium dehoogii are exceptional agents of disease. Three molecular methods targeting the partial β-tubulin gene were developed and evaluated to identify six closely related species of the S. apiospermum complex using quantitative real-time PCR (qPCR), PCR-based reverse line blot (PCR-RLB), and loop-mediated isothermal amplification (LAMP). qPCR was not specific enough for the identification of all species but had the highest sensitivity. The PCR-RLB assay was efficient for the identification of five species. LAMP distinguished all six species unambiguously. The analytical sensitivities of qPCR, PCR-RLB, and LAMP combined with MagNAPure, CTAB (cetyltrimethylammonium bromide), and FTA filter (Whatman) extraction were 50, 5 x 10³, and 5 x 10² cells/μl, respectively. When LAMP was combined with a simplified DNA extraction method using an FTA filter, identification to the species level was achieved within 2 h, including DNA extraction. The FTA-LAMP assay is therefore recommended as a cost-effective, simple, and rapid method for the identification of Scedosporium species. |
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Three molecular methods targeting the partial β-tubulin gene were developed and evaluated to identify six closely related species of the S. apiospermum complex using quantitative real-time PCR (qPCR), PCR-based reverse line blot (PCR-RLB), and loop-mediated isothermal amplification (LAMP). qPCR was not specific enough for the identification of all species but had the highest sensitivity. The PCR-RLB assay was efficient for the identification of five species. LAMP distinguished all six species unambiguously. The analytical sensitivities of qPCR, PCR-RLB, and LAMP combined with MagNAPure, CTAB (cetyltrimethylammonium bromide), and FTA filter (Whatman) extraction were 50, 5 x 10³, and 5 x 10² cells/μl, respectively. When LAMP was combined with a simplified DNA extraction method using an FTA filter, identification to the species level was achieved within 2 h, including DNA extraction. The FTA-LAMP assay is therefore recommended as a cost-effective, simple, and rapid method for the identification of Scedosporium species.</description><identifier>ISSN: 0095-1137</identifier><identifier>EISSN: 1098-660X</identifier><identifier>DOI: 10.1128/JCM.01813-10</identifier><identifier>PMID: 21177887</identifier><identifier>CODEN: JCMIDW</identifier><language>eng</language><publisher>Washington, DC: American Society for Microbiology</publisher><subject>Biological and medical sciences ; Fundamental and applied biological sciences. Psychology ; Fungal Proteins - genetics ; Humans ; Microbiology ; Molecular Diagnostic Techniques - methods ; Mycetoma - diagnosis ; Mycetoma - microbiology ; Mycology ; Mycology - methods ; Nucleic Acid Amplification Techniques - methods ; Pseudallescheria ; Pseudallescheria - genetics ; Pseudallescheria - isolation & purification ; Pseudallescheria boydii ; Scedosporium ; Scedosporium - genetics ; Scedosporium - isolation & purification ; Scedosporium apiospermum ; Scedosporium prolificans ; Sensitivity and Specificity ; Tubulin - genetics</subject><ispartof>Journal of Clinical Microbiology, 2011-03, Vol.49 (3), p.960-967</ispartof><rights>2015 INIST-CNRS</rights><rights>Copyright © 2011, American Society for Microbiology. 2011 American Society for Microbiology</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c494t-d5a23efa1a08bef06f676455ce3a21c73b057abc8b347b8c659a4c711bf77c883</citedby><cites>FETCH-LOGICAL-c494t-d5a23efa1a08bef06f676455ce3a21c73b057abc8b347b8c659a4c711bf77c883</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3067705/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3067705/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,3188,3189,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=23917439$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21177887$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Lu, Qiaoyun</creatorcontrib><creatorcontrib>Gerrits van den Ende, A.H.G</creatorcontrib><creatorcontrib>Bakkers, J.M.J.E</creatorcontrib><creatorcontrib>Sun, Jiufeng</creatorcontrib><creatorcontrib>Lackner, M</creatorcontrib><creatorcontrib>Najafzadeh, M.J</creatorcontrib><creatorcontrib>Melchers, W.J.G</creatorcontrib><creatorcontrib>Li, Ruoyu</creatorcontrib><creatorcontrib>de Hoog, G.S</creatorcontrib><title>Identification of Pseudallescheria and Scedosporium Species by Three Molecular Methods</title><title>Journal of Clinical Microbiology</title><addtitle>J Clin Microbiol</addtitle><description>The major clinically relevant species in Scedosporium (teleomorph Pseudallescheria) are Pseudallescheria boydii, Scedosporium aurantiacum, Scedosporium apiospermum, and Scedosporium prolificans, while Pseudallescheria minutispora, Petriellopsis desertorum, and Scedosporium dehoogii are exceptional agents of disease. Three molecular methods targeting the partial β-tubulin gene were developed and evaluated to identify six closely related species of the S. apiospermum complex using quantitative real-time PCR (qPCR), PCR-based reverse line blot (PCR-RLB), and loop-mediated isothermal amplification (LAMP). qPCR was not specific enough for the identification of all species but had the highest sensitivity. The PCR-RLB assay was efficient for the identification of five species. LAMP distinguished all six species unambiguously. The analytical sensitivities of qPCR, PCR-RLB, and LAMP combined with MagNAPure, CTAB (cetyltrimethylammonium bromide), and FTA filter (Whatman) extraction were 50, 5 x 10³, and 5 x 10² cells/μl, respectively. When LAMP was combined with a simplified DNA extraction method using an FTA filter, identification to the species level was achieved within 2 h, including DNA extraction. The FTA-LAMP assay is therefore recommended as a cost-effective, simple, and rapid method for the identification of Scedosporium species.</description><subject>Biological and medical sciences</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fungal Proteins - genetics</subject><subject>Humans</subject><subject>Microbiology</subject><subject>Molecular Diagnostic Techniques - methods</subject><subject>Mycetoma - diagnosis</subject><subject>Mycetoma - microbiology</subject><subject>Mycology</subject><subject>Mycology - methods</subject><subject>Nucleic Acid Amplification Techniques - methods</subject><subject>Pseudallescheria</subject><subject>Pseudallescheria - genetics</subject><subject>Pseudallescheria - isolation & purification</subject><subject>Pseudallescheria boydii</subject><subject>Scedosporium</subject><subject>Scedosporium - genetics</subject><subject>Scedosporium - isolation & purification</subject><subject>Scedosporium apiospermum</subject><subject>Scedosporium prolificans</subject><subject>Sensitivity and Specificity</subject><subject>Tubulin - genetics</subject><issn>0095-1137</issn><issn>1098-660X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><recordid>eNqF0UFv1DAQBeAIgehSuHGGXFAvpMzESWxfKqFVgaKuQNoWcbMmzmTjKokXewPqvydllwInTpbsT88zeknyHOEUMVdvPi5Xp4AKRYbwIFkgaJVVFXx9mCwAdJkhCnmUPInxBgCLoiwfJ0c5opRKyUXy5aLhcedaZ2nn_Jj6Nv0ceWqo7znajoOjlMYmXVtufNz64KYhXW_ZOo5pfZtedYE5Xfme7dRTSFe863wTnyaPWuojPzucx8n1u_Or5Yfs8tP7i-Xby8wWuthlTUm54JaQQNXcQtVWsppHtCwoRytFDaWk2qpaFLJWtio1FVYi1q2UVilxnJztc7dTPXBj510C9WYb3EDh1nhy5t-X0XVm478bAZWUUM4BJ4eA4L9NHHdmcNFy39PIfopGg8RCa1X9V6qyzEFgKWf5ei9t8DEGbu_nQTB3nZm5M_Ors_lm5i_-3uEe_y5pBq8OgKKlvg00Whf_OKFRFkLPLt27zm26Hy6woTiYGzuYQhthdHX318s9ackb2oQ55nqdAwpAXWgAJX4CQGa0TA</recordid><startdate>20110301</startdate><enddate>20110301</enddate><creator>Lu, Qiaoyun</creator><creator>Gerrits van den Ende, A.H.G</creator><creator>Bakkers, J.M.J.E</creator><creator>Sun, Jiufeng</creator><creator>Lackner, M</creator><creator>Najafzadeh, M.J</creator><creator>Melchers, W.J.G</creator><creator>Li, Ruoyu</creator><creator>de Hoog, G.S</creator><general>American Society for Microbiology</general><scope>FBQ</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>M7N</scope><scope>5PM</scope></search><sort><creationdate>20110301</creationdate><title>Identification of Pseudallescheria and Scedosporium Species by Three Molecular Methods</title><author>Lu, Qiaoyun ; Gerrits van den Ende, A.H.G ; Bakkers, J.M.J.E ; Sun, Jiufeng ; Lackner, M ; Najafzadeh, M.J ; Melchers, W.J.G ; Li, Ruoyu ; de Hoog, G.S</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c494t-d5a23efa1a08bef06f676455ce3a21c73b057abc8b347b8c659a4c711bf77c883</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Biological and medical sciences</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fungal Proteins - genetics</topic><topic>Humans</topic><topic>Microbiology</topic><topic>Molecular Diagnostic Techniques - methods</topic><topic>Mycetoma - diagnosis</topic><topic>Mycetoma - microbiology</topic><topic>Mycology</topic><topic>Mycology - methods</topic><topic>Nucleic Acid Amplification Techniques - methods</topic><topic>Pseudallescheria</topic><topic>Pseudallescheria - genetics</topic><topic>Pseudallescheria - isolation & purification</topic><topic>Pseudallescheria boydii</topic><topic>Scedosporium</topic><topic>Scedosporium - genetics</topic><topic>Scedosporium - isolation & purification</topic><topic>Scedosporium apiospermum</topic><topic>Scedosporium prolificans</topic><topic>Sensitivity and Specificity</topic><topic>Tubulin - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Lu, Qiaoyun</creatorcontrib><creatorcontrib>Gerrits van den Ende, A.H.G</creatorcontrib><creatorcontrib>Bakkers, J.M.J.E</creatorcontrib><creatorcontrib>Sun, Jiufeng</creatorcontrib><creatorcontrib>Lackner, M</creatorcontrib><creatorcontrib>Najafzadeh, M.J</creatorcontrib><creatorcontrib>Melchers, W.J.G</creatorcontrib><creatorcontrib>Li, Ruoyu</creatorcontrib><creatorcontrib>de Hoog, G.S</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Journal of Clinical Microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Lu, Qiaoyun</au><au>Gerrits van den Ende, A.H.G</au><au>Bakkers, J.M.J.E</au><au>Sun, Jiufeng</au><au>Lackner, M</au><au>Najafzadeh, M.J</au><au>Melchers, W.J.G</au><au>Li, Ruoyu</au><au>de Hoog, G.S</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Identification of Pseudallescheria and Scedosporium Species by Three Molecular Methods</atitle><jtitle>Journal of Clinical Microbiology</jtitle><addtitle>J Clin Microbiol</addtitle><date>2011-03-01</date><risdate>2011</risdate><volume>49</volume><issue>3</issue><spage>960</spage><epage>967</epage><pages>960-967</pages><issn>0095-1137</issn><eissn>1098-660X</eissn><coden>JCMIDW</coden><abstract>The major clinically relevant species in Scedosporium (teleomorph Pseudallescheria) are Pseudallescheria boydii, Scedosporium aurantiacum, Scedosporium apiospermum, and Scedosporium prolificans, while Pseudallescheria minutispora, Petriellopsis desertorum, and Scedosporium dehoogii are exceptional agents of disease. Three molecular methods targeting the partial β-tubulin gene were developed and evaluated to identify six closely related species of the S. apiospermum complex using quantitative real-time PCR (qPCR), PCR-based reverse line blot (PCR-RLB), and loop-mediated isothermal amplification (LAMP). qPCR was not specific enough for the identification of all species but had the highest sensitivity. The PCR-RLB assay was efficient for the identification of five species. LAMP distinguished all six species unambiguously. The analytical sensitivities of qPCR, PCR-RLB, and LAMP combined with MagNAPure, CTAB (cetyltrimethylammonium bromide), and FTA filter (Whatman) extraction were 50, 5 x 10³, and 5 x 10² cells/μl, respectively. When LAMP was combined with a simplified DNA extraction method using an FTA filter, identification to the species level was achieved within 2 h, including DNA extraction. The FTA-LAMP assay is therefore recommended as a cost-effective, simple, and rapid method for the identification of Scedosporium species.</abstract><cop>Washington, DC</cop><pub>American Society for Microbiology</pub><pmid>21177887</pmid><doi>10.1128/JCM.01813-10</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Biological and medical sciences Fundamental and applied biological sciences. Psychology Fungal Proteins - genetics Humans Microbiology Molecular Diagnostic Techniques - methods Mycetoma - diagnosis Mycetoma - microbiology Mycology Mycology - methods Nucleic Acid Amplification Techniques - methods Pseudallescheria Pseudallescheria - genetics Pseudallescheria - isolation & purification Pseudallescheria boydii Scedosporium Scedosporium - genetics Scedosporium - isolation & purification Scedosporium apiospermum Scedosporium prolificans Sensitivity and Specificity Tubulin - genetics |
title | Identification of Pseudallescheria and Scedosporium Species by Three Molecular Methods |
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