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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Development of a Real-Time PCR for a Sensitive One-Step Coprodiagnosis Allowing both the Identification of Carnivore Feces and the Detection of <named-content content-type="genus-species">Toxocara</named-content>
spp. and <named-content content-type="genus-species">Echinococcus multilocularis</named-content>
</title>
<author><name sortKey="Knapp, Jenny" sort="Knapp, Jenny" uniqKey="Knapp J" first="Jenny" last="Knapp">Jenny Knapp</name>
<affiliation><nlm:aff id="aff1">Chrono-environnement Laboratory, UMR UBFC/CNRS 6249 aff. INRA, University of Bourgogne Franche-Comté, Besançon, France</nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff2">Department of Parasitology-Mycology, University Hospital of Besançon, Besançon, France</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Umhang, Gerald" sort="Umhang, Gerald" uniqKey="Umhang G" first="Gérald" last="Umhang">Gérald Umhang</name>
<affiliation><nlm:aff id="aff3">ANSES Nancy Laboratory for Rabies and Wildlife, National Reference Laboratory for Echinococcus spp., Wildlife Surveillance and Eco-Epidemiology Unit, Technopole Agricole et Vétérinaire, Malzéville, France</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Poulle, Marie Lazarine" sort="Poulle, Marie Lazarine" uniqKey="Poulle M" first="Marie-Lazarine" last="Poulle">Marie-Lazarine Poulle</name>
<affiliation><nlm:aff id="aff4">University of Reims Champagne-Ardenne, SFR Cap-Santé, EA3800-PROTAL, UFR de Médecine, Reims, and CERFE, Boult-aux-Bois, France</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Millon, Laurence" sort="Millon, Laurence" uniqKey="Millon L" first="Laurence" last="Millon">Laurence Millon</name>
<affiliation><nlm:aff id="aff1">Chrono-environnement Laboratory, UMR UBFC/CNRS 6249 aff. INRA, University of Bourgogne Franche-Comté, Besançon, France</nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff2">Department of Parasitology-Mycology, University Hospital of Besançon, Besançon, France</nlm:aff>
</affiliation>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PMC</idno>
<idno type="pmid">26969697</idno>
<idno type="pmc">4959075</idno>
<idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4959075</idno>
<idno type="RBID">PMC:4959075</idno>
<idno type="doi">10.1128/AEM.03467-15</idno>
<date when="2016">2016</date>
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Development of a Real-Time PCR for a Sensitive One-Step Coprodiagnosis Allowing both the Identification of Carnivore Feces and the Detection of <named-content content-type="genus-species">Toxocara</named-content>
spp. and <named-content content-type="genus-species">Echinococcus multilocularis</named-content>
</title>
<author><name sortKey="Knapp, Jenny" sort="Knapp, Jenny" uniqKey="Knapp J" first="Jenny" last="Knapp">Jenny Knapp</name>
<affiliation><nlm:aff id="aff1">Chrono-environnement Laboratory, UMR UBFC/CNRS 6249 aff. INRA, University of Bourgogne Franche-Comté, Besançon, France</nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff2">Department of Parasitology-Mycology, University Hospital of Besançon, Besançon, France</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Umhang, Gerald" sort="Umhang, Gerald" uniqKey="Umhang G" first="Gérald" last="Umhang">Gérald Umhang</name>
<affiliation><nlm:aff id="aff3">ANSES Nancy Laboratory for Rabies and Wildlife, National Reference Laboratory for Echinococcus spp., Wildlife Surveillance and Eco-Epidemiology Unit, Technopole Agricole et Vétérinaire, Malzéville, France</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Poulle, Marie Lazarine" sort="Poulle, Marie Lazarine" uniqKey="Poulle M" first="Marie-Lazarine" last="Poulle">Marie-Lazarine Poulle</name>
<affiliation><nlm:aff id="aff4">University of Reims Champagne-Ardenne, SFR Cap-Santé, EA3800-PROTAL, UFR de Médecine, Reims, and CERFE, Boult-aux-Bois, France</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Millon, Laurence" sort="Millon, Laurence" uniqKey="Millon L" first="Laurence" last="Millon">Laurence Millon</name>
<affiliation><nlm:aff id="aff1">Chrono-environnement Laboratory, UMR UBFC/CNRS 6249 aff. INRA, University of Bourgogne Franche-Comté, Besançon, France</nlm:aff>
</affiliation>
<affiliation><nlm:aff id="aff2">Department of Parasitology-Mycology, University Hospital of Besançon, Besançon, France</nlm:aff>
</affiliation>
</author>
</analytic>
<series><title level="j">Applied and Environmental Microbiology</title>
<idno type="ISSN">0099-2240</idno>
<idno type="eISSN">1098-5336</idno>
<imprint><date when="2016">2016</date>
</imprint>
</series>
</biblStruct>
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<front><div type="abstract" xml:lang="en"><p>Studying the environmental occurrence of parasites of concern for humans and animals based on coprosamples is an expanding field of work in epidemiology and the ecology of health. Detecting and quantifying <named-content content-type="genus-species">Toxocara</named-content>
spp. and <named-content content-type="genus-species">Echinococcus multilocularis</named-content>
, two predominant zoonotic helminths circulating in European carnivores, in feces may help to better target measures for prevention. A rapid, sensitive, and one-step quantitative PCR (qPCR) allowing detection of <named-content content-type="genus-species">E. multilocularis</named-content>
and <named-content content-type="genus-species">Toxocara</named-content>
spp. was developed in the present study, combined with a host fecal test based on the identification of three carnivores (red fox, dog, and cat) involved in the life cycles of these parasites. A total of 68 coprosamples were collected from identified specimens from <named-content content-type="genus-species">Vulpes vulpes</named-content>
, <named-content content-type="genus-species">Canis lupus familiaris</named-content>
, <named-content content-type="genus-species">Canis lupus</named-content>
, <named-content content-type="genus-species">Felis silvestris catus</named-content>
, <named-content content-type="genus-species">Meles meles</named-content>
, <named-content content-type="genus-species">Martes foina</named-content>
, and <named-content content-type="genus-species">Martes martes</named-content>
. With DNA coprosamples, real-time PCR was performed in duplex with a qPCR inhibitor control specifically designed for this study. All the coprosample host identifications were confirmed by qPCR combined with sequencing, and parasites were detected and confirmed (<named-content content-type="genus-species">E. multilocularis</named-content>
in red foxes and <named-content content-type="genus-species">Toxocara cati</named-content>
in cats; 16% of samples presented inhibition). By combining parasite detection and quantification, the host fecal test, and a new qPCR inhibitor control, we created a technique with a high sensitivity that may considerably improve environmental studies of pathogens.</p>
</div>
</front>
</TEI>
<pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Appl Environ Microbiol</journal-id>
<journal-id journal-id-type="iso-abbrev">Appl. Environ. Microbiol</journal-id>
<journal-id journal-id-type="hwp">aem</journal-id>
<journal-id journal-id-type="pmc">aem</journal-id>
<journal-id journal-id-type="publisher-id">AEM</journal-id>
<journal-title-group><journal-title>Applied and Environmental Microbiology</journal-title>
</journal-title-group>
<issn pub-type="ppub">0099-2240</issn>
<issn pub-type="epub">1098-5336</issn>
<publisher><publisher-name>American Society for Microbiology</publisher-name>
<publisher-loc>1752 N St., N.W., Washington, DC</publisher-loc>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">26969697</article-id>
<article-id pub-id-type="pmc">4959075</article-id>
<article-id pub-id-type="publisher-id">03467-15</article-id>
<article-id pub-id-type="doi">10.1128/AEM.03467-15</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Genetics and Molecular Biology</subject>
</subj-group>
</article-categories>
<title-group><article-title>Development of a Real-Time PCR for a Sensitive One-Step Coprodiagnosis Allowing both the Identification of Carnivore Feces and the Detection of <named-content content-type="genus-species">Toxocara</named-content>
spp. and <named-content content-type="genus-species">Echinococcus multilocularis</named-content>
</article-title>
<alt-title alt-title-type="running-head"><named-content content-type="genus-species">Toxocara</named-content>
, <named-content content-type="genus-species">E. multilocularis</named-content>
, and Host Diagnosis</alt-title>
<alt-title alt-title-type="short-authors">Knapp et al.</alt-title>
</title-group>
<contrib-group><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid" authenticated="false">http://orcid.org/0000-0002-2054-1619</contrib-id>
<name><surname>Knapp</surname>
<given-names>Jenny</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>a</sup>
</xref>
<xref ref-type="aff" rid="aff2"><sup>b</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Umhang</surname>
<given-names>Gérald</given-names>
</name>
<xref ref-type="aff" rid="aff3"><sup>c</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Poulle</surname>
<given-names>Marie-Lazarine</given-names>
</name>
<xref ref-type="aff" rid="aff4"><sup>d</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Millon</surname>
<given-names>Laurence</given-names>
</name>
<xref ref-type="aff" rid="aff1"><sup>a</sup>
</xref>
<xref ref-type="aff" rid="aff2"><sup>b</sup>
</xref>
</contrib>
<aff id="aff1"><label>a</label>
Chrono-environnement Laboratory, UMR UBFC/CNRS 6249 aff. INRA, University of Bourgogne Franche-Comté, Besançon, France</aff>
<aff id="aff2"><label>b</label>
Department of Parasitology-Mycology, University Hospital of Besançon, Besançon, France</aff>
<aff id="aff3"><label>c</label>
ANSES Nancy Laboratory for Rabies and Wildlife, National Reference Laboratory for Echinococcus spp., Wildlife Surveillance and Eco-Epidemiology Unit, Technopole Agricole et Vétérinaire, Malzéville, France</aff>
<aff id="aff4"><label>d</label>
University of Reims Champagne-Ardenne, SFR Cap-Santé, EA3800-PROTAL, UFR de Médecine, Reims, and CERFE, Boult-aux-Bois, France</aff>
</contrib-group>
<contrib-group><contrib contrib-type="editor"><name><surname>Dozois</surname>
<given-names>C. M.</given-names>
</name>
<role>Editor</role>
</contrib>
<aff>INRS–Institut Armand-Frappier</aff>
</contrib-group>
<author-notes><corresp id="cor1">Address correspondence to Jenny Knapp, <email>jenny.knapp@univ-fcomte.fr</email>
.</corresp>
<fn fn-type="other"><p><bold>Citation</bold>
Knapp J, Umhang G, Poulle M-L, Millon L. 2016. Development of a real-time PCR for a sensitive one-step coprodiagnosis allowing both the identification of carnivore feces and the detection of <named-content content-type="genus-species">Toxocara</named-content>
spp. and <named-content content-type="genus-species">Echinococcus multilocularis</named-content>
. Appl Environ Microbiol 82:2950–2958. doi:<ext-link ext-link-type="uri" xlink:href="http://dx.doi.org/10.1128/AEM.03467-15">10.1128/AEM.03467-15</ext-link>
.</p>
</fn>
</author-notes>
<pub-date pub-type="epreprint"><day>11</day>
<month>3</month>
<year>2016</year>
</pub-date>
<pub-date pub-type="epub"><day>2</day>
<month>5</month>
<year>2016</year>
</pub-date>
<pub-date pub-type="collection"><day>15</day>
<month>5</month>
<year>2016</year>
</pub-date>
<volume>82</volume>
<issue>10</issue>
<fpage>2950</fpage>
<lpage>2958</lpage>
<history><date date-type="received"><day>22</day>
<month>10</month>
<year>2015</year>
</date>
<date date-type="accepted"><day>3</day>
<month>3</month>
<year>2016</year>
</date>
</history>
<permissions><copyright-statement>Copyright © 2016, American Society for Microbiology. All Rights Reserved.</copyright-statement>
<copyright-year>2016</copyright-year>
<copyright-holder>American Society for Microbiology</copyright-holder>
</permissions>
<self-uri content-type="pdf" xlink:href="zam01016002950.pdf"></self-uri>
<abstract><p>Studying the environmental occurrence of parasites of concern for humans and animals based on coprosamples is an expanding field of work in epidemiology and the ecology of health. Detecting and quantifying <named-content content-type="genus-species">Toxocara</named-content>
spp. and <named-content content-type="genus-species">Echinococcus multilocularis</named-content>
, two predominant zoonotic helminths circulating in European carnivores, in feces may help to better target measures for prevention. A rapid, sensitive, and one-step quantitative PCR (qPCR) allowing detection of <named-content content-type="genus-species">E. multilocularis</named-content>
and <named-content content-type="genus-species">Toxocara</named-content>
spp. was developed in the present study, combined with a host fecal test based on the identification of three carnivores (red fox, dog, and cat) involved in the life cycles of these parasites. A total of 68 coprosamples were collected from identified specimens from <named-content content-type="genus-species">Vulpes vulpes</named-content>
, <named-content content-type="genus-species">Canis lupus familiaris</named-content>
, <named-content content-type="genus-species">Canis lupus</named-content>
, <named-content content-type="genus-species">Felis silvestris catus</named-content>
, <named-content content-type="genus-species">Meles meles</named-content>
, <named-content content-type="genus-species">Martes foina</named-content>
, and <named-content content-type="genus-species">Martes martes</named-content>
. With DNA coprosamples, real-time PCR was performed in duplex with a qPCR inhibitor control specifically designed for this study. All the coprosample host identifications were confirmed by qPCR combined with sequencing, and parasites were detected and confirmed (<named-content content-type="genus-species">E. multilocularis</named-content>
in red foxes and <named-content content-type="genus-species">Toxocara cati</named-content>
in cats; 16% of samples presented inhibition). By combining parasite detection and quantification, the host fecal test, and a new qPCR inhibitor control, we created a technique with a high sensitivity that may considerably improve environmental studies of pathogens.</p>
</abstract>
<counts><fig-count count="2"></fig-count>
<table-count count="6"></table-count>
<equation-count count="0"></equation-count>
<ref-count count="30"></ref-count>
<page-count count="9"></page-count>
<word-count count="7903"></word-count>
</counts>
</article-meta>
</front>
</pmc>
</record>
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