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Open AccessArticle

Genotyping Echinococcus multilocularis in Human Alveolar Echinococcosis Patients: An EmsB Microsatellite Analysis

1
UMR CNRS 6249 Laboratoire Chrono-environnement, Université Franche-Comté, 16 Route de Gray, 25030 Besançon, France
2
Department of Parasitology-Mycology, National Reference Centre for Echinococcoses, University Hospital of Besançon, 25030 Besançon, France
3
Institute for Infectious Diseases, Faculty of Medicine, University of Berne, 3001 Berne, Switzerland
4
Parasitology-Mycology Laboratory, Lariboisière-Saint Louis-Fernand Widal Hospital, Assistance Publique-Hôpitaux de Paris (AP-HP), Université de Paris, 75475 Paris, France
5
UMR INTERTRYP, IRD/CIRAD, University of Montpellier, 34398 Montpellier, France
6
ANSES, Nancy Laboratory for Rabies and Wildlife, Wildlife Surveillance and Eco-Epidemiology Unit, Technopôle Agricole et Vétérinaire, B.P. 40009, 54220 Malzéville, France
7
Department of Hepatology, University Hospital of Besançon, 25000 Besançon, France
*
Author to whom correspondence should be addressed.
Pathogens 2020, 9(4), 282; https://doi.org/10.3390/pathogens9040282
Received: 9 March 2020 / Revised: 6 April 2020 / Accepted: 8 April 2020 / Published: 13 April 2020
(This article belongs to the Special Issue Echinococcus)
For clinical epidemiology specialists, connecting the genetic diversity of Echinococcus multilocularis to sources of infection or particular sites has become somewhat of a holy grail. It is very difficult to trace the infection history of alveolar echinococcosis (AE) patients as there may be an incubation period of five to 15 years before reliable diagnosis. Moreover, the variability of parasitic manifestations in human patients raises the possibility of genetically different isolates of E. multilocularis having different levels of pathogenicity. Thus, the exposure of human patients to different strains or genotypes circulating in geographically different environments may lead to different disease outcomes. Molecular tools, such as the microsatellite marker EmsB, were required to investigate these aspects. This genetic marker was previously tested on a collection of 1211 European field samples predominantly of animal origin, referenced on a publicly available database. In this study, we investigated a panel of 66 metacestode samples (between 1981 and 2019) recovered surgically from 63 patients diagnosed with alveolar echinococcosis originating from four European countries (France, Switzerland, Germany, Belgium). In this study, we identified nine EmsB profiles, five of which were found in patients located in the same areas of France and Switzerland. One profile was detected on both sides of the French-Swiss border, whereas most patients from non-endemic regions clustered together in another profile. EmsB profiles appeared to remain stable over time because similar profiles were detected in patients who underwent surgery recently and patients who underwent surgery some time ago. This study sheds light on possible pathways of contamination in humans, including proximity contamination in some cases, and the dominant contamination profiles in Europe, particularly for extrahepatic lesions. View Full-Text
Keywords: Alveolar echinococcosis; Echinococcus multilocularis; European endemic area; contamination event; genotyping; microsatellite EmsB Alveolar echinococcosis; Echinococcus multilocularis; European endemic area; contamination event; genotyping; microsatellite EmsB
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    Doi: 10.5281/zenodo.3701246
    Link: http://doi.org/10.5281/zenodo.3701246
    Description: Supplementary Materials: Table S1: Euclidean distance matrix for 5 patients tested (1HP: 1-HP-102039-BE-Bru ; BON: BON-363-FR-39 ; 15HP : 15-HP-061502-GE-Bw ; 42HP: 42-HP-122345-SW-Fr), for the MLX-A to D and PL PCR conditions and FSA Table S2: Euclidean distance matrix for 66 isolates tested in MLX-A and PL PCR conditions and by FSA Table S3: Euclidean distance matrix for the 14 EchinoRisk worms tested (PAHO5-117 to 120 from a German fox, UCPR1-495 to 499 from a Czech fox, and 16PL-375 to 379 from a Polish fox) from the IPA, in MLX-A and PL conditions and by FSA Table S4: Individual similarities between the 66 tested isolates and EWET reference data, and by country of origin Figure S1: Dendrogram constructed from EmsB amplification data obtained under MLX-A and PL PCR conditions and by FSA for the 66 patient isolates tested, based on hierarchical clustering analysis (Euclidean distance and unweighted pair group method).
MDPI and ACS Style

Knapp, J.; Gottstein, B.; Bretagne, S.; Bart, J.-M.; Umhang, G.; Richou, C.; Bresson-Hadni, S.; Millon, L. Genotyping Echinococcus multilocularis in Human Alveolar Echinococcosis Patients: An EmsB Microsatellite Analysis. Pathogens 2020, 9, 282.

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