title: Evaluation by latent class analysis of a magnetic capture based DNA extraction followed by real-time qPCR as a new diagnostic method for detection of Echinococcus multilocularis in definitive hosts.
abstract: A new method, based on a magnetic capture based DNA extraction followed by qPCR, was developed for the detection of the zoonotic parasite Echinococcus multilocularis in definitive hosts. Latent class analysis was used to compare this new method with the currently used phenol-chloroform DNA extraction followed by single tube nested PCR. In total, 60 red foxes and coyotes from three different locations were tested with both molecular methods and the sedimentation and counting technique (SCT) or intestinal scraping technique (IST). Though based on a limited number of samples, it could be established that the magnetic capture based DNA extraction followed by qPCR showed similar sensitivity and specificity as the currently used phenol-chloroform DNA extraction followed by single tube nested PCR. All methods have a high specificity as shown by Bayesian latent class analysis. Both molecular assays have higher sensitivities than the combined SCT and IST, though the uncertainties in sensitivity estimates were wide for all assays tested. The magnetic capture based DNA extraction followed by qPCR has the advantage of not requiring hazardous chemicals like the phenol-chloroform DNA extraction followed by single tube nested PCR. This supports the replacement of the phenol-chloroform DNA extraction followed by single tube nested PCR by the magnetic capture based DNA extraction followed by qPCR for molecular detection of E. multilocularis in definitive hosts.
authors.affiliation: Centre for Infectious Disease Control, National Institute for Public Health and the Environment (RIVM), 3720 BA Bilthoven, The Netherlands. Electronic address: miriam.maas@rivm.nlvan.
authors.forename: Miriam
authors.initials: M
authors.lastname: Maas
authors.affiliation: Centre for Infectious Disease Control, National Institute for Public Health and the Environment (RIVM), 3720 BA Bilthoven, The Netherlands.
authors.forename: Annika
authors.initials: A
authors.lastname: van Roon
authors.affiliation: Centre for Infectious Disease Control, National Institute for Public Health and the Environment (RIVM), 3720 BA Bilthoven, The Netherlands.
authors.forename: Cecile
authors.initials: C
authors.lastname: Dam-Deisz
authors.affiliation: Centre for Infectious Disease Control, National Institute for Public Health and the Environment (RIVM), 3720 BA Bilthoven, The Netherlands.
authors.forename: Marieke
authors.initials: M
authors.lastname: Opsteegh
authors.affiliation: Wildlife Ecology and Spatial Epidemiology Lab (WEaSEL), Department of Ecosystem and Public Health, Faculty of Veterinary Medicine, University of Calgary, 3280 Hospital Drive, NW, Calgary, AB, T2N 4Z6, Canada.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Massolo
authors.affiliation: Institute of Food Safety, Animal Health and Environment "BIOR", Lejupes Street 3, LV-1076 Riga, Latvia.
authors.forename: Gunita
authors.initials: G
authors.lastname: Deksne
authors.affiliation: Centre for Infectious Disease Control, National Institute for Public Health and the Environment (RIVM), 3720 BA Bilthoven, The Netherlands.
authors.forename: Peter
authors.initials: P
authors.lastname: Teunis
authors.affiliation: Centre for Infectious Disease Control, National Institute for Public Health and the Environment (RIVM), 3720 BA Bilthoven, The Netherlands.
authors.forename: Joke
authors.initials: J
authors.lastname: van der Giessen
mesh_terms: D000818:Animals; D002170:Canada; D046029:Coyotes / Q000469:parasitology; D018509:DNA, Helminth / Q000235:genetics / Q000302:isolation & purification; D004443:Echinococcosis / Q000175:diagnosis / Q000662:veterinary*; D048210:Echinococcus multilocularis / Q000235:genetics / Q000502:physiology*; D005243:Feces / Q000469:parasitology; D005589:Foxes / Q000469:parasitology; D007844:Latvia; D008280:Magnetics; D009426:Netherlands; D010274:Parasitology / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction; D012680:Sensitivity and Specificity
chemical_list: D018509:DNA, Helminth
keywords: Coyotes; Echinococcus multilocularis; Latent class analysis; Magnetic capture based DNA extraction; Real-time qPCR; Red foxes