A semi-automated magnetic capture probe based DNA extraction and real-time PCR method applied in the Swedish surveillance of Echinococcus multilocularis in red fox (Vulpes vulpes) faecal samples
Identifieur interne : 000130 ( Main/Exploration ); précédent : 000129; suivant : 000131A semi-automated magnetic capture probe based DNA extraction and real-time PCR method applied in the Swedish surveillance of Echinococcus multilocularis in red fox (Vulpes vulpes) faecal samples
Auteurs : Mats Isaksson [Suède] ; Sa Hagström [Suède] ; Maria Teresa Armua-Fernandez [Suisse] ; Helene Wahlström [Suède] ; Erik Olof Gren [Suède] ; Andrea Miller [Suède] ; Anders Holmberg [Suède] ; Morten Lukacs [Suède] ; Adriano Casulli [Italie] ; Peter Deplazes [Suisse] ; Mikael Juremalm [Suède]Source :
- Parasites & Vectors [ 1756-3305 ] ; 2014.
Abstract
Following the first finding of
We describe a new semi-automated magnetic capture probe DNA extraction method and real time hydrolysis probe polymerase chain reaction assay (MC-PCR) for the detection of
Of 177 foxes analysed by the sedimentation and counting technique,
The diagnostic specificity of the MC-PCR was evaluated using fox scats collected within the Swedish screening. Of 2158 samples analysed, two were positive. This implies that the specificity is at least 99.9% (C.I. = 99.7 -100).
The MC-PCR proved to have a high sensitivity and a very high specificity. The test is partially automated but also possible to perform manually if desired. The test is well suited for nationwide
Url:
DOI: 10.1186/s13071-014-0583-6
PubMed: 25522844
PubMed Central: 4282741
Affiliations:
- Italie, Suisse, Suède
- Canton de Zurich, Latium, Svealand
- Rome, Stockholm, Zurich
- Université de Zurich
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Le document en format XML
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type="region">Canton de Zurich</region></placeName></affiliation></author><author><name sortKey="Wahlstrom, Helene" sort="Wahlstrom, Helene" uniqKey="Wahlstrom H" first="Helene" last="Wahlström">Helene Wahlström</name><affiliation wicri:level="1"><nlm:aff id="Aff3">Department of Epidemiology, National Veterinary Institute, Uppsala, Sweden</nlm:aff><country xml:lang="fr">Suède</country><wicri:regionArea>Department of Epidemiology, National Veterinary Institute, Uppsala</wicri:regionArea><wicri:noRegion>Uppsala</wicri:noRegion></affiliation></author><author><name sortKey=" Gren, Erik Olof" sort=" Gren, Erik Olof" uniqKey=" Gren E" first="Erik Olof" last=" Gren">Erik Olof Gren</name><affiliation wicri:level="1"><nlm:aff id="Aff4">Department of Pathology and Wildlife Diseases, National Veterinary Institute, Uppsala, Sweden</nlm:aff><country xml:lang="fr">Suède</country><wicri:regionArea>Department of Pathology and Wildlife Diseases, National Veterinary Institute, Uppsala</wicri:regionArea><wicri:noRegion>Uppsala</wicri:noRegion></affiliation></author><author><name sortKey="Miller, Andrea" sort="Miller, Andrea" uniqKey="Miller A" first="Andrea" last="Miller">Andrea Miller</name><affiliation wicri:level="1"><nlm:aff id="Aff5">Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, Uppsala, Sweden</nlm:aff><country xml:lang="fr">Suède</country><wicri:regionArea>Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, Uppsala</wicri:regionArea><wicri:noRegion>Uppsala</wicri:noRegion></affiliation></author><author><name sortKey="Holmberg, Anders" sort="Holmberg, Anders" uniqKey="Holmberg A" first="Anders" last="Holmberg">Anders Holmberg</name><affiliation wicri:level="3"><nlm:aff id="Aff6">Precision System Science, Stockholm, Sweden</nlm:aff><country xml:lang="fr">Suède</country><wicri:regionArea>Precision System Science, 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Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanità, Rome</wicri:regionArea><placeName><settlement type="city">Rome</settlement><region nuts="2">Latium</region></placeName></affiliation></author><author><name sortKey="Deplazes, Peter" sort="Deplazes, Peter" uniqKey="Deplazes P" first="Peter" last="Deplazes">Peter Deplazes</name><affiliation wicri:level="4"><nlm:aff id="Aff2">Institute of Parasitology, Vetsuisse and Medical Faculty, University of Zurich, Zurich, Switzerland</nlm:aff><country xml:lang="fr">Suisse</country><wicri:regionArea>Institute of Parasitology, Vetsuisse and Medical Faculty, University of Zurich, Zurich</wicri:regionArea><orgName type="university">Université de Zurich</orgName><placeName><settlement type="city">Zurich</settlement><region nuts="3" type="region">Canton de Zurich</region></placeName></affiliation></author><author><name sortKey="Juremalm, Mikael" sort="Juremalm, Mikael" uniqKey="Juremalm M" first="Mikael" last="Juremalm">Mikael Juremalm</name><affiliation wicri:level="1"><nlm:aff id="Aff1">Department of Virology Immunobiology and Parasitology, National Veterinary Institute, Uppsala, Sweden</nlm:aff><country xml:lang="fr">Suède</country><wicri:regionArea>Department of Virology Immunobiology and Parasitology, National Veterinary Institute, Uppsala</wicri:regionArea><wicri:noRegion>Uppsala</wicri:noRegion></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">25522844</idno><idno type="pmc">4282741</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4282741</idno><idno type="RBID">PMC:4282741</idno><idno type="doi">10.1186/s13071-014-0583-6</idno><date when="2014">2014</date><idno type="wicri:Area/Pmc/Corpus">000016</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000016</idno><idno type="wicri:Area/Pmc/Curation">000016</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000016</idno><idno type="wicri:Area/Pmc/Checkpoint">000044</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">000044</idno><idno type="wicri:Area/Ncbi/Merge">000394</idno><idno type="wicri:Area/Ncbi/Curation">000394</idno><idno type="wicri:Area/Ncbi/Checkpoint">000394</idno><idno type="wicri:Area/Main/Merge">000130</idno><idno type="wicri:Area/Main/Curation">000130</idno><idno type="wicri:Area/Main/Exploration">000130</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">A semi-automated magnetic capture probe based DNA extraction and real-time PCR method applied in the Swedish surveillance of <italic>Echinococcus multilocularis</italic> in red fox (<italic>Vulpes vulpes</italic>) faecal samples</title><author><name sortKey="Isaksson, Mats" sort="Isaksson, Mats" uniqKey="Isaksson M" first="Mats" last="Isaksson">Mats Isaksson</name><affiliation wicri:level="1"><nlm:aff id="Aff1">Department of Virology Immunobiology and Parasitology, National Veterinary Institute, Uppsala, Sweden</nlm:aff><country xml:lang="fr">Suède</country><wicri:regionArea>Department of Virology Immunobiology and Parasitology, National Veterinary Institute, Uppsala</wicri:regionArea><wicri:noRegion>Uppsala</wicri:noRegion></affiliation></author><author><name sortKey="Hagstrom, Sa" sort="Hagstrom, Sa" uniqKey="Hagstrom " first=" Sa" last="Hagström"> Sa Hagström</name><affiliation wicri:level="1"><nlm:aff id="Aff1">Department of Virology Immunobiology and Parasitology, National Veterinary Institute, Uppsala, Sweden</nlm:aff><country xml:lang="fr">Suède</country><wicri:regionArea>Department of Virology Immunobiology and Parasitology, National Veterinary Institute, Uppsala</wicri:regionArea><wicri:noRegion>Uppsala</wicri:noRegion></affiliation></author><author><name sortKey="Armua Fernandez, Maria Teresa" sort="Armua Fernandez, Maria Teresa" uniqKey="Armua Fernandez M" first="Maria Teresa" last="Armua-Fernandez">Maria Teresa Armua-Fernandez</name><affiliation wicri:level="4"><nlm:aff id="Aff2">Institute of Parasitology, Vetsuisse and Medical Faculty, University of Zurich, Zurich, Switzerland</nlm:aff><country xml:lang="fr">Suisse</country><wicri:regionArea>Institute of Parasitology, Vetsuisse and Medical Faculty, University of Zurich, Zurich</wicri:regionArea><orgName type="university">Université de Zurich</orgName><placeName><settlement type="city">Zurich</settlement><region nuts="3" type="region">Canton de Zurich</region></placeName></affiliation></author><author><name sortKey="Wahlstrom, Helene" sort="Wahlstrom, Helene" uniqKey="Wahlstrom H" first="Helene" last="Wahlström">Helene Wahlström</name><affiliation wicri:level="1"><nlm:aff id="Aff3">Department of Epidemiology, National Veterinary Institute, Uppsala, Sweden</nlm:aff><country xml:lang="fr">Suède</country><wicri:regionArea>Department of Epidemiology, National Veterinary Institute, Uppsala</wicri:regionArea><wicri:noRegion>Uppsala</wicri:noRegion></affiliation></author><author><name sortKey=" Gren, Erik Olof" sort=" Gren, Erik Olof" uniqKey=" Gren E" first="Erik Olof" last=" Gren">Erik Olof Gren</name><affiliation wicri:level="1"><nlm:aff id="Aff4">Department of Pathology and Wildlife Diseases, National Veterinary Institute, Uppsala, Sweden</nlm:aff><country xml:lang="fr">Suède</country><wicri:regionArea>Department of Pathology and Wildlife Diseases, National Veterinary Institute, Uppsala</wicri:regionArea><wicri:noRegion>Uppsala</wicri:noRegion></affiliation></author><author><name sortKey="Miller, Andrea" sort="Miller, Andrea" uniqKey="Miller A" first="Andrea" last="Miller">Andrea Miller</name><affiliation wicri:level="1"><nlm:aff id="Aff5">Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, Uppsala, Sweden</nlm:aff><country xml:lang="fr">Suède</country><wicri:regionArea>Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, Uppsala</wicri:regionArea><wicri:noRegion>Uppsala</wicri:noRegion></affiliation></author><author><name sortKey="Holmberg, Anders" sort="Holmberg, Anders" uniqKey="Holmberg A" first="Anders" last="Holmberg">Anders Holmberg</name><affiliation wicri:level="3"><nlm:aff id="Aff6">Precision System Science, Stockholm, Sweden</nlm:aff><country xml:lang="fr">Suède</country><wicri:regionArea>Precision System Science, Stockholm</wicri:regionArea><placeName><settlement type="city">Stockholm</settlement><region nuts="2">Svealand</region></placeName></affiliation></author><author><name sortKey="Lukacs, Morten" sort="Lukacs, Morten" uniqKey="Lukacs M" first="Morten" last="Lukacs">Morten Lukacs</name><affiliation wicri:level="3"><nlm:aff id="Aff6">Precision System Science, Stockholm, Sweden</nlm:aff><country xml:lang="fr">Suède</country><wicri:regionArea>Precision System Science, Stockholm</wicri:regionArea><placeName><settlement type="city">Stockholm</settlement><region nuts="2">Svealand</region></placeName></affiliation></author><author><name sortKey="Casulli, Adriano" sort="Casulli, Adriano" uniqKey="Casulli A" first="Adriano" last="Casulli">Adriano Casulli</name><affiliation wicri:level="3"><nlm:aff id="Aff7">Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanità, Rome, Italy</nlm:aff><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanità, Rome</wicri:regionArea><placeName><settlement type="city">Rome</settlement><region nuts="2">Latium</region></placeName></affiliation></author><author><name sortKey="Deplazes, Peter" sort="Deplazes, Peter" uniqKey="Deplazes P" first="Peter" last="Deplazes">Peter Deplazes</name><affiliation wicri:level="4"><nlm:aff id="Aff2">Institute of Parasitology, Vetsuisse and Medical Faculty, University of Zurich, Zurich, Switzerland</nlm:aff><country xml:lang="fr">Suisse</country><wicri:regionArea>Institute of Parasitology, Vetsuisse and Medical Faculty, University of Zurich, Zurich</wicri:regionArea><orgName type="university">Université de Zurich</orgName><placeName><settlement type="city">Zurich</settlement><region nuts="3" type="region">Canton de Zurich</region></placeName></affiliation></author><author><name sortKey="Juremalm, Mikael" sort="Juremalm, Mikael" uniqKey="Juremalm M" first="Mikael" last="Juremalm">Mikael Juremalm</name><affiliation wicri:level="1"><nlm:aff id="Aff1">Department of Virology Immunobiology and Parasitology, National Veterinary Institute, Uppsala, Sweden</nlm:aff><country xml:lang="fr">Suède</country><wicri:regionArea>Department of Virology Immunobiology and Parasitology, National Veterinary Institute, Uppsala</wicri:regionArea><wicri:noRegion>Uppsala</wicri:noRegion></affiliation></author></analytic><series><title level="j">Parasites & Vectors</title><idno type="eISSN">1756-3305</idno><imprint><date when="2014">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><sec><title>Background</title><p>Following the first finding of <italic>Echinococcus multilocularis</italic> in Sweden in 2011, 2985 red foxes (<italic>Vulpes vulpes</italic>) were analysed by the segmental sedimentation and counting technique. This is a labour intensive method and requires handling of the whole carcass of the fox, resulting in a costly analysis. In an effort to reduce the cost of labour and sample handling, an alternative method has been developed. The method is sensitive and partially automated for detection of <italic>E. multilocularis</italic> in faecal samples. The method has been used in the Swedish <italic>E. multilocularis</italic> monitoring program for 2012–2013 on more than 2000 faecal samples.</p></sec><sec><title>Methods</title><p>We describe a new semi-automated magnetic capture probe DNA extraction method and real time hydrolysis probe polymerase chain reaction assay (MC-PCR) for the detection of <italic>E. multilocularis</italic> DNA in faecal samples from red fox. The diagnostic sensitivity was determined by validating the new method against the sedimentation and counting technique in fox samples collected in Switzerland where <italic>E. multilocularis</italic> is highly endemic.</p></sec><sec><title>Results</title><p>Of 177 foxes analysed by the sedimentation and counting technique, <italic>E. multilocularis</italic> was detected in 93 animals. Eighty-two (88%, 95% C.I 79.8-93.9) of these were positive in the MC-PCR. In foxes with more than 100 worms, the MC-PCR was positive in 44 out of 46 (95.7%) cases. The two MC-PCR negative samples originated from foxes with only immature <italic>E. multilocularis</italic> worms. In foxes with 100 worms or less, (<italic>n</italic> = 47), 38 (80.9%) were positive in the MC-PCR.</p><p>The diagnostic specificity of the MC-PCR was evaluated using fox scats collected within the Swedish screening. Of 2158 samples analysed, two were positive. This implies that the specificity is at least 99.9% (C.I. = 99.7 -100).</p></sec><sec><title>Conclusions</title><p>The MC-PCR proved to have a high sensitivity and a very high specificity. The test is partially automated but also possible to perform manually if desired. The test is well suited for nationwide <italic>E. multilocularis</italic> surveillance programs where sampling of fox scats is done to reduce the costs for sampling and where a test with a high sensitivity and a very high specificity is needed.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct></biblStruct><biblStruct></biblStruct><biblStruct><analytic><author><name sortKey="Umhang, G" uniqKey="Umhang G">G Umhang</name></author><author><name sortKey="Woronoff Rhen, N" uniqKey="Woronoff Rhen N">N Woronoff-Rhen</name></author><author><name sortKey="Combes, B" uniqKey="Combes B">B Combes</name></author><author><name sortKey="Boue, F" uniqKey="Boue F">F Boue</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bretagne, S" uniqKey="Bretagne S">S Bretagne</name></author><author><name sortKey="Guillou, Jp" uniqKey="Guillou J">JP Guillou</name></author><author><name sortKey="Morand, M" uniqKey="Morand M">M 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J" uniqKey="Huggett J">J Huggett</name></author><author><name sortKey="Kubista, M" uniqKey="Kubista M">M Kubista</name></author><author><name sortKey="Mueller, R" uniqKey="Mueller R">R Mueller</name></author><author><name sortKey="Nolan, T" uniqKey="Nolan T">T Nolan</name></author><author><name sortKey="Pfaffl, Mw" uniqKey="Pfaffl M">MW Pfaffl</name></author><author><name sortKey="Shipley, Gl" uniqKey="Shipley G">GL Shipley</name></author><author><name sortKey="Vandesompele, J" uniqKey="Vandesompele J">J Vandesompele</name></author><author><name sortKey="Wittwer, Ct" uniqKey="Wittwer C">CT Wittwer</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Monnier, P" uniqKey="Monnier P">P Monnier</name></author><author><name sortKey="Cliquet, F" uniqKey="Cliquet F">F Cliquet</name></author><author><name sortKey="Aubert, M" uniqKey="Aubert M">M Aubert</name></author><author><name sortKey="Bretagne, S" uniqKey="Bretagne S">S Bretagne</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Al Sabi, Mn" uniqKey="Al Sabi M">MN Al-Sabi</name></author><author><name sortKey="Kapel, Cm" uniqKey="Kapel C">CM Kapel</name></author><author><name sortKey="Deplazes, P" uniqKey="Deplazes P">P Deplazes</name></author><author><name sortKey="Mathis, A" uniqKey="Mathis A">A Mathis</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Oines, O" uniqKey="Oines O">O Oines</name></author><author><name sortKey="Isaksson, M" uniqKey="Isaksson M">M Isaksson</name></author><author><name sortKey="Hagstrom, A" uniqKey="Hagstrom A">A Hagstrom</name></author><author><name sortKey="Tavornpanich, S" uniqKey="Tavornpanich S">S Tavornpanich</name></author><author><name sortKey="Davidson, Rk" uniqKey="Davidson R">RK Davidson</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Braasch, D" uniqKey="Braasch D">D Braasch</name></author><author><name sortKey="Corey, D" uniqKey="Corey D">D Corey</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kok, J" uniqKey="Kok J">J Kok</name></author><author><name sortKey="Wiegerinck, E" uniqKey="Wiegerinck E">E Wiegerinck</name></author><author><name sortKey="Giesendorf, B" uniqKey="Giesendorf B">B Giesendorf</name></author><author><name sortKey="Swinkels, D" uniqKey="Swinkels D">D Swinkels</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kapel, Cm" uniqKey="Kapel C">CM Kapel</name></author><author><name sortKey="Torgerson, Pr" uniqKey="Torgerson P">PR Torgerson</name></author><author><name sortKey="Thompson, Rc" uniqKey="Thompson R">RC Thompson</name></author><author><name sortKey="Deplazes, P" uniqKey="Deplazes P">P Deplazes</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Burlet, P" uniqKey="Burlet P">P Burlet</name></author><author><name sortKey="Deplazes, P" uniqKey="Deplazes P">P Deplazes</name></author><author><name sortKey="Hegglin, D" uniqKey="Hegglin D">D Hegglin</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>Italie</li><li>Suisse</li><li>Suède</li></country><region><li>Canton de Zurich</li><li>Latium</li><li>Svealand</li></region><settlement><li>Rome</li><li>Stockholm</li><li>Zurich</li></settlement><orgName><li>Université de Zurich</li></orgName></list><tree><country name="Suède"><noRegion><name sortKey="Isaksson, Mats" sort="Isaksson, Mats" uniqKey="Isaksson M" first="Mats" last="Isaksson">Mats Isaksson</name></noRegion><name sortKey=" Gren, Erik Olof" sort=" Gren, Erik Olof" uniqKey=" Gren E" first="Erik Olof" last=" Gren">Erik Olof Gren</name><name sortKey="Hagstrom, Sa" sort="Hagstrom, Sa" uniqKey="Hagstrom " first=" Sa" last="Hagström"> Sa Hagström</name><name sortKey="Holmberg, Anders" sort="Holmberg, Anders" uniqKey="Holmberg A" first="Anders" last="Holmberg">Anders Holmberg</name><name sortKey="Juremalm, Mikael" sort="Juremalm, Mikael" uniqKey="Juremalm M" first="Mikael" last="Juremalm">Mikael Juremalm</name><name sortKey="Lukacs, Morten" sort="Lukacs, Morten" uniqKey="Lukacs M" first="Morten" last="Lukacs">Morten Lukacs</name><name sortKey="Miller, Andrea" sort="Miller, Andrea" uniqKey="Miller A" first="Andrea" last="Miller">Andrea Miller</name><name sortKey="Wahlstrom, Helene" sort="Wahlstrom, Helene" uniqKey="Wahlstrom H" first="Helene" last="Wahlström">Helene Wahlström</name></country><country name="Suisse"><region name="Canton de Zurich"><name sortKey="Armua Fernandez, Maria Teresa" sort="Armua Fernandez, Maria Teresa" uniqKey="Armua Fernandez M" first="Maria Teresa" last="Armua-Fernandez">Maria Teresa Armua-Fernandez</name></region><name sortKey="Deplazes, Peter" sort="Deplazes, Peter" uniqKey="Deplazes P" first="Peter" last="Deplazes">Peter Deplazes</name></country><country name="Italie"><region name="Latium"><name sortKey="Casulli, Adriano" sort="Casulli, Adriano" uniqKey="Casulli A" first="Adriano" last="Casulli">Adriano Casulli</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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