caa a22 4500 445341629 CHVBK 20180317142815.0 cr unu---uuuuu 170323e20110801xx s 000 0 eng 10.1007/s00436-011-2272-0 doi (NATIONALLICENCE)springer-10.1007/s00436-011-2272-0 A real-time multiplex-nested PCR system for coprological diagnosis of Echinococcus multilocularis and host species [Elektronische Daten] [Anke Dinkel, Selina Kern, Anja Brinker, Rainer Oehme, Amélie Vaniscotte, Patrick Giraudoux, Ute Mackenstedt, Thomas Romig] A hybridization probe-based real-time multiplex-nested PCR system was developed for the simultaneous detection of Echinococcus multilocularis and host species directly from faecal samples. Species identification was determined by melting curve analysis. Specificity was assessed by using DNA extracted from various cestodes (E. multilocularis, Echinococcus granulosus (G1), Echinococcus ortleppi, Echinococcus canadensis (G6, G7), Taenia crassiceps, Taenia hydatigena, Taenia mustelae, Taenia pisiformis, Taenia serialis, Taenia taeniaeformis, Mesocestoides leptothylacus), carnivores (Vulpes vulpes, Vulpes corsac, Vulpes ferrilata, Canis familiaris, Felis catus, Martes foina), Microtus arvalis and Arvicola terrestris. The analytical sensitivity was 10fg, evaluated with serially diluted DNA of E. multilocularis to 10μl total DNA solution from E. multilocularis-negative canid faeces. Based on a comparison of 47 dog samples from China, the proportion of the E. multilocularis-positive-tested samples by the real-time multiplex-nested PCR was moderately higher (38% vs. 30%) as when tested with a previously evaluated nested PCR with a sensitivity of 70-100%, depending on the number and gravidity status of worms present in the intestine (Dinkel et al., J Clin Microbiol 36:1871-1876, 1998). To assess the epidemiological applicability of this method, 227 canid faecal samples collected in the field were analysed. This newly developed real-time multiplex-nested PCR system is a specific, sensitive and reliable method for the detection of E. multilocularis and host species in faecal samples for epidemiological purposes. Springer-Verlag, 2011 Dinkel Anke Department of Parasitology, University of Hohenheim, Emil-Wolff-Str.34, 70599, Stuttgart, Germany aut Kern Selina Department of Parasitology, University of Hohenheim, Emil-Wolff-Str.34, 70599, Stuttgart, Germany aut Brinker Anja Department of Parasitology, University of Hohenheim, Emil-Wolff-Str.34, 70599, Stuttgart, Germany aut Oehme Rainer State Health Office Baden-Wuerttemberg, Nordbahnhofstr. 135, 70191, Stuttgart, Germany aut Vaniscotte Amélie Department of Chrono-environment, UMR UFC/CNRS 6249 aff. INRA, Universite de Franche-Comté, 25030, Besançon, France aut Giraudoux Patrick Department of Chrono-environment, UMR UFC/CNRS 6249 aff. INRA, Universite de Franche-Comté, 25030, Besançon, France aut Mackenstedt Ute Department of Parasitology, University of Hohenheim, Emil-Wolff-Str.34, 70599, Stuttgart, Germany aut Romig Thomas Department of Parasitology, University of Hohenheim, Emil-Wolff-Str.34, 70599, Stuttgart, Germany aut Parasitology Research Springer-Verlag 109/2(2011-08-01), 493-498 0932-0113 109:2<493 2011 109 436 https://doi.org/10.1007/s00436-011-2272-0 text/html Onlinezugriff via DOI 1 brief-communication jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00436-011-2272-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Dinkel Anke Department of Parasitology, University of Hohenheim, Emil-Wolff-Str.34, 70599, Stuttgart, Germany aut NATIONALLICENCE 700 1- Kern Selina Department of Parasitology, University of Hohenheim, Emil-Wolff-Str.34, 70599, Stuttgart, Germany aut NATIONALLICENCE 700 1- Brinker Anja Department of Parasitology, University of Hohenheim, Emil-Wolff-Str.34, 70599, Stuttgart, Germany aut NATIONALLICENCE 700 1- Oehme Rainer State Health Office Baden-Wuerttemberg, Nordbahnhofstr. 135, 70191, Stuttgart, Germany aut NATIONALLICENCE 700 1- Vaniscotte Amélie Department of Chrono-environment, UMR UFC/CNRS 6249 aff. INRA, Universite de Franche-Comté, 25030, Besançon, France aut NATIONALLICENCE 700 1- Giraudoux Patrick Department of Chrono-environment, UMR UFC/CNRS 6249 aff. INRA, Universite de Franche-Comté, 25030, Besançon, France aut NATIONALLICENCE 700 1- Mackenstedt Ute Department of Parasitology, University of Hohenheim, Emil-Wolff-Str.34, 70599, Stuttgart, Germany aut NATIONALLICENCE 700 1- Romig Thomas Department of Parasitology, University of Hohenheim, Emil-Wolff-Str.34, 70599, Stuttgart, Germany aut NATIONALLICENCE 773 0- Parasitology Research Springer-Verlag 109/2(2011-08-01), 493-498 0932-0113 109:2<493 2011 109 436 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer