Real time PCR to detect the environmental faecal contamination by Echinococcus multilocularis from red fox stools.

• Published in: Veterinary Parasitology
• Main Authors: Knapp, Jenny, Millon, Laurence, Mouzon, Lorane, Umhang, Gérald, Raoul, Francis, Ali, Zeinaba Said, Combes, Benoît, Comte, Sébastien, Gbaguidi-Haore, Houssein, Grenouillet, Frédéric, Giraudoux, Patrick
• Other Authors: Laboratoire Chrono-environnement (UMR 6249) (LCE), Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté COMUE (UBFC)-Université Bourgogne Franche-Comté COMUE (UBFC), Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon), Laboratoire de la rage et pathologie des animaux sauvages (LERPAS), Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Entente de Lutte Interdépartementale contre les Zoonoses (ELIZ), Etablissement Public Interdépartemental, Institut universitaire de France (IUF), Ministère de l'Education nationale, de l’Enseignement supérieur et de la Recherche (M.E.N.E.S.R.), JSPS/CNRS funding
• Format: Article in Journal/Newspaper
• Language: English
• Published: HAL CCSD 2014
• Subjects: Echinococcus multilocularis; SSCT; POLYMERASE-CHAIN-REACTION; EGGS; CARNIVORES; FECES; Vulpes vulpes; Environmental contamination; DNA; qPCR; Fox faeces; VULPES-VULPES; DIAGNOSIS; LIMITS; [SDV]Life Sciences [q-bio]; Greenland; Jura
• Online Access: https://hal.science/hal-00943686; https://doi.org/10.1016/j.vetpar.2013.12.023

International audience : The oncosphere stage of Echinococcus multilocularis in red fox stools can lead, after ingestion, to the development of alveolar echinococcosis in the intermediate hosts, commonly small mammals and occasionally humans. Monitoring animal infection and environmental contamination is a key issue in public health surveillance. We developed a quantitative real-time PCR technique (qPCR) to detect and quantify E. multilocularis DNA released in fox faeces. A qPCR technique using a hydrolysis probe targeting part of the mitochondrial gene rrnL was assessed on (i) a reference collection of stools from 57 necropsied foxes simultaneously investigated using the segmental sedimentation and counting technique (SSCT) (29 positive for E. multilocularis worms and 28 negative animals for the parasite); (ii) a collection of 114 fox stools sampled in the field: two sets of 50 samples from contrasted endemic regions in France and 14 from an E. multilocularis-free area (Greenland). Of the negative SSCT controls, 26/28 were qPCR-negative and two were weakly positive. Of the positive SSCT foxes, 25/29 samples were found to be positive by qPCR. Of the field samples, qPCR was positive in 21/50 (42%) and 5/48 (10.4%) stools (2 samples inhibited), originating respectively from high and low endemic areas. In faeces, averages of 0.1pg/μl of DNA in the Jura area and 0.7pg/μl in the Saône-et-Loire area were detected. All qPCR-positive samples were confirmed by sequencing. The qPCR technique developed here allowed us to quantify environmental E. multilocularis contamination by fox faeces by studying the infectious agent directly. No previous study had performed this test in a one-step reaction.