title: Development of a Real-Time PCR for a Sensitive One-Step Coprodiagnosis Allowing both the Identification of Carnivore Feces and the Detection of Toxocara spp. and Echinococcus multilocularis.
abstract: 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 Toxocara spp. and Echinococcus multilocularis, 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 E. multilocularis and Toxocara 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 Vulpes vulpes, Canis lupus familiaris, Canis lupus, Felis silvestris catus, Meles meles, Martes foina, and Martes martes 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 (E. multilocularis in red foxes and Toxocara cati 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.
authors.affiliation: Chrono-environnement Laboratory, UMR UBFC/CNRS 6249 aff. INRA, University of Bourgogne Franche-Comté, Besançon, France jenny.knapp@univ-fcomte.fr.|Department of Parasitology-Mycology, University Hospital of Besançon, Besançon, France.
authors.forename: Jenny
authors.identifier: http://orcid.org/0000-0002-2054-1619
authors.initials: J
authors.lastname: Knapp
authors.affiliation: 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.
authors.forename: Gérald
authors.initials: G
authors.lastname: Umhang
authors.affiliation: University of Reims Champagne-Ardenne, SFR Cap-Santé, EA3800-PROTAL, UFR de Médecine, Reims, and CERFE, Boult-aux-Bois, France.
authors.forename: Marie-Lazarine
authors.initials: ML
authors.lastname: Poulle
authors.affiliation: Chrono-environnement Laboratory, UMR UBFC/CNRS 6249 aff. INRA, University of Bourgogne Franche-Comté, Besançon, France.|Department of Parasitology-Mycology, University Hospital of Besançon, Besançon, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Millon
mesh_terms: D000818:Animals; D002415:Cats; D004285:Dogs; D048210:Echinococcus multilocularis / Q000235:genetics / Q000302:isolation & purification*; D005243:Feces / Q000469:parasitology*; D005589:Foxes; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014119:Toxocara / Q000235:genetics / Q000302:isolation & purification*