Hostname: page-component-cd9895bd7-lnqnp Total loading time: 0 Render date: 2024-12-25T01:10:50.787Z Has data issue: false hasContentIssue false
  • English
  • Français

Simple modification to improve reliability of copro-DNA examinations for diagnosing Echinococcus multilocularis infections in red foxes

Published online by Cambridge University Press:  14 April 2020

T. Irie Open the ORCID record for T. Irie [Opens in a new window] ,
T. Ito ,
H. Kouguchi  and
K. Uraguchi
T. Irie*
Affiliation:
Medical Zoology Group, Department of Infectious Diseases, Hokkaido Institute of Public Health, North 19, West 12, Kitaku, Sapporo, Hokkaido060-0819, Japan
T. Ito
Affiliation:
Medical Zoology Group, Department of Infectious Diseases, Hokkaido Institute of Public Health, North 19, West 12, Kitaku, Sapporo, Hokkaido060-0819, Japan
H. Kouguchi
Affiliation:
Medical Zoology Group, Department of Infectious Diseases, Hokkaido Institute of Public Health, North 19, West 12, Kitaku, Sapporo, Hokkaido060-0819, Japan
K. Uraguchi
Affiliation:
Medical Zoology Group, Department of Infectious Diseases, Hokkaido Institute of Public Health, North 19, West 12, Kitaku, Sapporo, Hokkaido060-0819, Japan
*
Author for correspondence: T. Irie, E-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Abstract

Epidemiological studies of Echinococcus multilocularis infections in definitive hosts require a reliable and economic diagnostic method. In this study, the current copro-DNA examination technique was modified by increasing the faecal amounts tested and adding a step to neutralize the faeces before DNA extraction. Reliability of the modified method was evaluated using rectal faecal samples from red foxes and comparing them with intestinal worms detected using the sedimentation and counting technique (SCT) following necropsy. The modified copro-DNA examination method demonstrated 93.9% sensitivity (138/147) on the SCT. Its detectability increased depending on the worm burden, and the sensitivity was 100% in cases harbouring over 1000 worms. From 111 SCT-negative cases, six (5.4%) were copro-DNA-positive, and all were confirmed as E. multilocularis via sequencing analysis. Five of the remaining 105 SCT-negative cases (4.8%) retained polymerase chain reaction (PCR) inhibitors in the extracted solution, suggesting that approximately 5% of the red fox faeces retained these inhibitors after treatment with the present copro-DNA extraction method. Although further evaluation is needed for faeces deposited in the wild, the present copro-DNA examination technique will help monitor the E. multilocularis prevalence in definitive hosts. When used for detailed evaluations of endemicity (e.g. changes in infection pressure or spread in non-endemic areas), the absence of PCR inhibitors should be confirmed, and multiple trials on faecal subsamples are recommended.

Keywords

Copro-DNAEchinococcus multilocularis infectionintestinal wormred fox
Type
Short Communication
Information
Journal of Helminthology , Volume 94 , 2020 , e145
Copyright
Copyright © The Author(s), 2020. Published by Cambridge University Press

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Conraths, FJ and Deplazes, P (2015) Echinococcus multilocularis: epidemiology, surveillance and state-of-the-art diagnostics from a veterinary public health perspective. Veterinary Parasitology 213, 149–161.CrossRefGoogle ScholarPubMed
Deplazes, P and Eckert, J (2001) Veterinary aspects of alveolar echinococcosis-a zoonosis of public health significance. Veterinary Parasitology 98, 65–87.CrossRefGoogle ScholarPubMed
Dinkel, A, von Nickisch-Rosenegk, M, Bilger, B, Merli, M, Lucius, R and Romig, T (1998) Detection of Echinococcus multilocularis in the definitive host: coprodiagnosis by PCR as an alternative to necropsy. Journal of Clinical Microbiology 36, 1871–1876.CrossRefGoogle ScholarPubMed
Dinkel, A, Kern, S, Brinker, A, Oehme, R, Vaniscotte, A, Giraudoux, P, Mackenstedt, U and Romig, T (2011) A real-time multiplex-nested PCR system for coprological diagnosis of Echinococcus multilocularis and host species. Parasitology Research 109, 493–498.CrossRefGoogle ScholarPubMed
Eckert, J, Deplazes, P, Craig, PS, Gemmell, MA, Gottstein, B, Heath, D, Jenkins, DJ, Kamiya, M and Lightowlers, M (2001) Echinococcosis in animals: clinical aspects, diagnosis and treatment. pp. 72–100in Eckert, J, Gemmell, MA, Meslin, FX and Pawlowski, ZS (Eds) WHO/OIE manual on echinococcosis in humans and animals: a public health problem of global concern. Paris, France, WHO/OIE.Google Scholar
Hofer, S, Gloor, S, Müller, U, Mathis, A, Hegglin, D and Deplazes, P (2000) High prevalence of Echinococcus multilocularis in urban red foxes (Vulpes vulpes) and voles (Arvicola terrestris) in the city of Zurich, Switzerland. Parasitology 120, 135–142.CrossRefGoogle ScholarPubMed
Irie, T, Ito, T, Kouguchi, H, Yamano, K, Uraguchi, K, Yagi, K and Nonaka, N (2017) Diagnosis of canine Echinococcus multilocularis infections by copro-DNA tests: comparison of DNA extraction techniques and evaluation of diagnostic deworming. Parasitology Research 116, 2139–2144.CrossRefGoogle ScholarPubMed
Isaksson, M, Hagstrom, A, Armua-Fernandez, MT, et al. (2014) A semiautomated magnetic capture probe based DNA extraction and real-time PCR method applied in the Swedish surveillance of Echinococcus multilocularis in red fox (Vulpes vulpes) faecal samples. Parasites & Vectors 7, 583.CrossRefGoogle Scholar
Kohn, M and Wayne, RK (1997) Facts from feces revisited. Trends in Ecology & Evolution 12, 223–227.CrossRefGoogle ScholarPubMed
König, A, Romig, T and Holzhofer, E (2019) Effective long-term control of Echinococcus multilocularis in a mixed rural-urban area in southern Germany. PLoS ONE 14, e0214993.CrossRefGoogle Scholar
Maas, M, van Roon, A, Dam-Deisz, C, Opsteegh, M, Massolo, A, Deksne, G, Teunis, P and van der Giessen, J (2016) 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. Veterinary Parasitology 230, 20–24.CrossRefGoogle ScholarPubMed
Maksimov, P, Schares, G, Press, S, Fröhlich, A, Basso, W, Herzig, M and Conraths, FJ (2017) Comparison of different commercial DNA extraction kits and PCR protocols for the detection of Echinococcus multilocularis eggs in faecal samples from foxes. Veterinary Parasitology 237, 83–93.CrossRefGoogle ScholarPubMed
Maksimov, P, Isaksson, M, Schares, G, Romig, T and Conraths, FJ (2019) Validation of PCR-based protocols for the detection of Echinococcus multilocularis DNA in the final host using the intestinal scraping technique as a reference. Food and Waterborne Parasitology 15, e00044.CrossRefGoogle ScholarPubMed
Monteiro, L, Bonnemaison, D, Vekris, A, Petry, KG, Bonnet, J, Vidal, R, Cabrita, J and Mégraud, F (1997) Complex polysaccharides as PCR inhibitors in feces: Helicobacter pylori model. Journal of Clinical Microbiology 35, 995–998.Google ScholarPubMed
Monteiro, L, Gras, N and Megraud, F (2001) Magnetic immuno-PCR assay with inhibitor removal for direct detection of Helicobacter pylori in human feces. Journal of Clinical Microbiology 39, 3778–3780.CrossRefGoogle ScholarPubMed
Nonaka, N, Iida, M, Yagi, K, Ito, T, Ooi, HK, Oku, Y and Kamiya, M (1996) Time course of coproantigen excretion in Echinococcus multilocularis infections in foxes and an alternative definitive host, golden hamsters. International Journal for Parasitology 26, 1271–1278.CrossRefGoogle Scholar
Tackmann, K, Mattis, R and Conraths, FJ (2006) Detection of Echinococcus multilocularis in foxes: Evaluation of a protocol of the intestinal scraping technique. Journal of Veterinary Medicine. B, Infectious Diseases and Veterinary Public Health 53, 395–398.CrossRefGoogle ScholarPubMed
Supplementary material: File

Irie et al. supplementary material

Irie et al. supplementary material

Download Irie et al. supplementary material(File)
File 39.9 KB