{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,5]],"date-time":"2026-09-05T21:51:23Z","timestamp":1788645083167,"version":"build-2803163510"},"reference-count":35,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2020,9,26]],"date-time":"2020-09-26T00:00:00Z","timestamp":1601078400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100010661","name":"Horizon 2020 Framework Programme","doi-asserted-by":"publisher","award":["773830"],"award-info":[{"award-number":["773830"]}],"id":[{"id":"10.13039\/100010661","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Infections with eggs of Echinococcus granulosus sensu lato (s.l.) can cause cystic echinococcosis in intermediate host animals and humans. Upon ingestion of viable eggs, oncospheres hatch from the eggs and subsequently develop into fluid-filled larval cysts, most frequently in the liver or the lungs. The slowly growing cysts progressively interfere with organ function. The risk of infection is determined by the host range of the parasite, its pathogenicity and other epidemiologically relevant parameters, which differ significantly among the five species within the E. granulosus s.l. complex. It is therefore essential to diagnose the correct species within E. granulosus s.l. to help understand specific disease epidemiology and to facilitate effective implementation of control measures. For this purpose, simple, fast and cost-effective typing techniques are needed. We developed quantitative real-time polymerase chain reactions (qPCRs) to target polymorphic regions in the mitochondrial genome of E. granulosus s.l. In a single-step typing approach, we distinguished E. granulosus s.l. members in four epidemiologically relevant subgroups. These were E. granulosus sensu stricto, E. equinus, E. ortleppi and the E. canadensis cluster. The technique also allowed identification and differentiation of these species from other Echinococcus or Taenia taxa for samples isolated from cysts or faeces.<\/jats:p>","DOI":"10.3390\/pathogens9100791","type":"journal-article","created":{"date-parts":[[2020,9,27]],"date-time":"2020-09-27T22:24:42Z","timestamp":1601245482000},"page":"791","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":28,"title":["Species Detection within the Echinococcus granulosus sensu lato Complex by Novel Probe-Based Real-Time PCRs"],"prefix":"10.3390","volume":"9","author":[{"given":"Pavlo","family":"Maksimov","sequence":"first","affiliation":[{"name":"Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Epidemiology, National Reference Centre for Echinococcosis, 17493 Greifswald-Insel Riems, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3487-7241","authenticated-orcid":false,"given":"Hannes","family":"Bergmann","sequence":"additional","affiliation":[{"name":"Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Epidemiology, National Reference Centre for Echinococcosis, 17493 Greifswald-Insel Riems, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marion","family":"Wassermann","sequence":"additional","affiliation":[{"name":"Parasitology Unit, University of Hohenheim, Emil-Wolff-Str. 34, 70599 Stuttgart, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Romig","sequence":"additional","affiliation":[{"name":"Parasitology Unit, University of Hohenheim, Emil-Wolff-Str. 34, 70599 Stuttgart, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0782-3723","authenticated-orcid":false,"given":"Bruno","family":"Gottstein","sequence":"additional","affiliation":[{"name":"Institute of Parasitology, Vetsuisse Faculty and Faculty of Medicine, University of Bern, 3012 Bern, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9183-7591","authenticated-orcid":false,"given":"Adriano","family":"Casulli","sequence":"additional","affiliation":[{"name":"WHO Collaborating Centre for the Epidemiology, Detection and Control of Cystic and Alveolar Echinococcosis, Department of Infectious Diseases, Istituto Superiore di Sanit\u00e0, 00161 Rome, Italy"},{"name":"European Reference Laboratory for Parasites, Department of Infectious Diseases, Istituto Superiore di Sanit\u00e0, 00161 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7400-9409","authenticated-orcid":false,"given":"Franz J.","family":"Conraths","sequence":"additional","affiliation":[{"name":"Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Institute of Epidemiology, National Reference Centre for Echinococcosis, 17493 Greifswald-Insel Riems, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,9,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1051\/parasite\/2020024","article-title":"World Association of Echinococcosis International consensus on terminology to be used in the field of echinococcoses","volume":"27","author":"Vuitton","year":"2020","journal-title":"Parasite"},{"key":"ref_2","first-page":"2088","article-title":"Risk factors for alveolar echinococcosis in humans","volume":"10","author":"Kern","year":"2004","journal-title":"Emerg. Infect. Dis."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Bristow, B.N., Lee, S., Shafir, S., and Sorvillo, F. (2012). Human echinococcosis mortality in the United States, 1990-2007. PLoS Negl. Trop. Dis., 6.","DOI":"10.1371\/journal.pntd.0001524"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1016\/j.vetpar.2015.07.035","article-title":"Taxonomy and molecular epidemiology of Echinococcus granulosus sensu lato","volume":"213","author":"Romig","year":"2015","journal-title":"Vet. Parasitol"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"292","DOI":"10.3201\/eid1402.070742","article-title":"Emergence of polycystic neotropical echinococcosis","volume":"14","author":"Tappe","year":"2008","journal-title":"Emerg. Infect. Dis."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1016\/bs.apar.2016.11.002","article-title":"Ecology and Life Cycle Patterns of Echinococcus Species","volume":"95","author":"Romig","year":"2017","journal-title":"Adv. Parasitol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1016\/j.meegid.2017.04.023","article-title":"New mitogenome and nuclear evidence on the phylogeny and taxonomy of the highly zoonotic tapeworm Echinococcus granulosus sensu stricto","volume":"52","author":"Kinkar","year":"2017","journal-title":"Infect Genet Evol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"923","DOI":"10.1016\/j.ijpara.2017.06.003","article-title":"Genetic differentiation of the G6\/7 cluster of Echinococcus canadensis based on mitochondrial marker genes","volume":"47","author":"Addy","year":"2017","journal-title":"Int. J. Parasitol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/j.ijppaw.2014.12.002","article-title":"A sylvatic lifecycle of Echinococcus equinus in the Etosha National Park, Namibia","volume":"4","author":"Wassermann","year":"2015","journal-title":"Int. J. Parasitol. Parasites Wildl."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1017","DOI":"10.1016\/j.ijpara.2013.06.002","article-title":"Phylogenetic systematics of the genus Echinococcus (Cestoda: Taeniidae)","volume":"43","author":"Nakao","year":"2013","journal-title":"Int. J. Parasitol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1625","DOI":"10.1017\/S0031182013000565","article-title":"Mitochondrial phylogeny of the genus Echinococcus (Cestoda: Taeniidae) with emphasis on relationships among Echinococcus canadensis genotypes","volume":"140","author":"Nakao","year":"2013","journal-title":"Parasitology"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"444","DOI":"10.1016\/j.meegid.2010.01.011","article-title":"State-of-the-art Echinococcus and Taenia: Phylogenetic taxonomy of human-pathogenic tapeworms and its application to molecular diagnosis","volume":"10","author":"Nakao","year":"2010","journal-title":"Infect. Genet. Evol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1016\/j.vetpar.2015.07.028","article-title":"Echinococcus granulosus: Epidemiology and state-of-the-art of diagnostics in animals","volume":"213","author":"Craig","year":"2015","journal-title":"Vet. Parasitol"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1016\/bs.apar.2016.09.003","article-title":"Laboratory Diagnosis of Echinococcus spp. in Human Patients and Infected Animals","volume":"96","author":"Casulli","year":"2017","journal-title":"Adv. Parasitol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1111\/j.1439-0450.2006.01003.x","article-title":"Detection of Echinococcus multilocularis in foxes: Evaluation of a protocol of the intestinal scraping technique","volume":"53","author":"Tackmann","year":"2006","journal-title":"J. Vet. Med. B. Infect. Dis. Vet. Public Health"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1016\/j.exppara.2011.01.004","article-title":"Segmental sedimentation and counting technique (SSCT): An adaptable method for qualitative diagnosis of Echinococcus multilocularis in fox intestines","volume":"128","author":"Umhang","year":"2011","journal-title":"Exp. Parasitol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1016\/S0001-706X(02)00216-4","article-title":"Predictive values and quality control of techniques for the diagnosis of Echinococcus multilocularis in definitive hosts","volume":"85","author":"Eckert","year":"2003","journal-title":"Acta Trop."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1016\/j.vetpar.2015.07.027","article-title":"Echinococcus multilocularis: Epidemiology, surveillance and state-of-the-art diagnostics from a veterinary public health perspective","volume":"213","author":"Conraths","year":"2015","journal-title":"Vet. Parasitol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1007\/s00436-004-1260-z","article-title":"Scraping or shaking--a comparison of methods for the quantitative determination of Echinococcus multilocularis in fox intestines","volume":"95","author":"Duscher","year":"2005","journal-title":"Parasitol. Res."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"911","DOI":"10.1017\/S0031182007002235","article-title":"Identification of taeniid eggs in the faeces from carnivores based on multiplex PCR using targets in mitochondrial DNA","volume":"134","author":"Trachsel","year":"2007","journal-title":"Parasitology"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Boubaker, G., Macchiaroli, N., Prada, L., Cucher, M.A., Rosenzvit, M.C., Ziadinov, I., Deplazes, P., Saarma, U., Babba, H., and Gottstein, B. (2013). A multiplex PCR for the simultaneous detection and genotyping of the Echinococcus granulosus complex. PLoS Negl. Trop. Dis., 7.","DOI":"10.1371\/journal.pntd.0002017"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/j.vetpar.2013.12.023","article-title":"Real time PCR to detect the environmental faecal contamination by Echinococcus multilocularis from red fox stools","volume":"201","author":"Knapp","year":"2014","journal-title":"Vet. Parasitol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"246","DOI":"10.1186\/1756-3305-7-246","article-title":"Laboratory assessment of sensitive molecular tools for detection of low levels of Echinococcus multilocularis-eggs in fox (Vulpes vulpes) faeces","volume":"7","author":"Oines","year":"2014","journal-title":"Parasit. Vectors"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"200","DOI":"10.1016\/j.jviromet.2006.05.020","article-title":"A universal heterologous internal control system for duplex real-time RT-PCR assays used in a detection system for pestiviruses","volume":"136","author":"Hoffmann","year":"2006","journal-title":"J. Virol. Methods"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"e470","DOI":"10.1016\/S2214-109X(20)30066-8","article-title":"Recognising the substantial burden of neglected pandemics cystic and alveolar echinococcosis","volume":"8","author":"Casulli","year":"2020","journal-title":"Lancet Glob. Health"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"296","DOI":"10.3201\/eid1202.050499","article-title":"Global socioeconomic impact of cystic echinococcosis","volume":"12","author":"Budke","year":"2006","journal-title":"Emerg. Infect. Dis."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"663","DOI":"10.1016\/j.ijpara.2016.04.005","article-title":"A novel zoonotic genotype related to Echinococcus granulosus sensu stricto from southern Ethiopia","volume":"46","author":"Wassermann","year":"2016","journal-title":"Int. J. Parasitol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1470","DOI":"10.1373\/clinchem.2013.203653","article-title":"Single-nucleotide polymorphisms and other mismatches reduce performance of quantitative PCR assays","volume":"59","author":"Lefever","year":"2013","journal-title":"Clin. Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1929","DOI":"10.1017\/S0031182018000719","article-title":"Molecular phylogeny based on six nuclear genes suggests that Echinococcus granulosus sensu lato genotypes G6\/G7 and G8\/G10 can be regarded as two distinct species","volume":"145","author":"Laurimae","year":"2018","journal-title":"Parasitology"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1016\/j.vetpar.2017.02.015","article-title":"Comparison of different commercial DNA extraction kits and PCR protocols for the detection of Echinococcus multilocularis eggs in faecal samples from foxes","volume":"237","author":"Maksimov","year":"2017","journal-title":"Vet. Parasitol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"113","DOI":"10.3354\/dao02053","article-title":"BSA reduces inhibition in a TaqMan assay for the detection of Batrachochytrium dendrobatidis","volume":"92","author":"Garland","year":"2010","journal-title":"Dis. Aquat. Organ."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"631","DOI":"10.1017\/S0031182002001749","article-title":"Spatial and temporal aspects of urban transmission of Echinococcus multilocularis","volume":"124","author":"Stieger","year":"2002","journal-title":"Parasitology"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"e00044","DOI":"10.1016\/j.fawpar.2019.e00044","article-title":"Validation of PCR-based protocols for the detection of Echinococcus multilocularis DNA in the final host using the Intestinal Scraping Technique as a reference","volume":"15","author":"Maksimov","year":"2019","journal-title":"Food Waterborne Parasitol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1016\/j.bdq.2015.01.005","article-title":"How good is a PCR efficiency estimate: Recommendations for precise and robust qPCR efficiency assessments","volume":"3","author":"Svec","year":"2015","journal-title":"Biomol. Detect. Quantif."},{"key":"ref_35","unstructured":"Scientific, T.F. (2020, September 25). DNA Copy Number and Dilution Calculator. Available online: https:\/\/www.thermofisher.com\/de\/de\/home\/brands\/thermo-scientific\/molecular-biology\/molecular-biology-learning-center\/molecular-biology-resource-library\/thermo-scientific-web-tools\/dna-copy-number-calculator.html."}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/9\/10\/791\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:13:54Z","timestamp":1760177634000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/9\/10\/791"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,9,26]]},"references-count":35,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2020,10]]}},"alternative-id":["pathogens9100791"],"URL":"https:\/\/doi.org\/10.3390\/pathogens9100791","relation":{"has-preprint":[{"id-type":"doi","id":"10.21203\/rs.3.rs-57339\/v1","asserted-by":"object"},{"id-type":"doi","id":"10.1101\/2020.07.24.220756","asserted-by":"object"}]},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,9,26]]}}}