Development of a real-time PCR for a sensitive one-step copro-diagnosis allowing both the identification of carnivore feces and the detection of Toxocara spp. and Echinococcus multilocularis.

• Published in: Applied and Environmental Microbiology
• Main Authors: Knapp, Jenny, Umhang, Gérald, Poulle, Marie-Lazarine, Millon, Laurence
• 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), Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Université de Reims Champagne-Ardenne (URCA), Projet Interdisciplinaire Ecologie de la Sante grant from the CNRS-INEE; InVS National Reference Center for Alveolar Echinococcosis
• Format: Article in Journal/Newspaper
• Language: English
• Published: CCSD 2016
• Subjects: [SDV.EE.SANT]Life Sciences [q-bio]/Ecology; environment/Health; Canis lupus
• Online Access: https://hal.science/hal-01294614; https://doi.org/10.1128/AEM.03467-15

International audience 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.