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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
medicine
A Multiplex PCR for the Simultaneous Detection and Genotyping of the Echinococcus granulosus Complex
PLoS Neglected Tropical Diseases, Volume 7, No. 1, Article e2017, Year 2013
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Description
Echinococcus granulosus is characterized by high intra-specific variability (genotypes G1-G10) and according to the new molecular phylogeny of the genus Echinococcus, the E. granulosus complex has been divided into E. granulosus sensu stricto (G1-G3), E. equinus (G4), E. ortleppi (G5), and E. canadensis (G6-G10). The molecular characterization of E. granulosus isolates is fundamental to understand the spatio-temporal epidemiology of this complex in many endemic areas with the simultaneous occurrence of different Echinococcus species and genotypes. To simplify the genotyping of the E. granulosus complex we developed a single-tube multiplex PCR (mPCR) allowing three levels of discrimination: (i) Echinococcus genus, (ii) E. granulosus complex in common, and (iii) the specific genotype within the E. granulosus complex. The methodology was established with known DNA samples of the different strains/genotypes, confirmed on 42 already genotyped samples (Spain: 22 and Bulgaria: 20) and then successfully applied on 153 unknown samples (Tunisia: 114, Algeria: 26 and Argentina: 13). The sensitivity threshold of the mPCR was found to be 5 ng Echinoccoccus DNA in a mixture of up to 1 μg of foreign DNA and the specificity was 100% when template DNA from closely related members of the genus Taenia was used. Additionally to DNA samples, the mPCR can be carried out directly on boiled hydatid fluid or on alkaline-lysed frozen or fixed protoscoleces, thus avoiding classical DNA extractions. However, when using Echinococcus eggs obtained from fecal samples of infected dogs, the sensitivity of the mPCR was low (<40%). Thus, except for copro analysis, the mPCR described here has a high potential for a worldwide application in large-scale molecular epidemiological studies on the Echinococcus genus. © 2013 Boubaker et al.
Available Materials
https://efashare.b-cdn.net/share/pmc/articles/PMC3547860/bin/pntd.0002017.s001.docx
https://efashare.b-cdn.net/share/pmc/articles/PMC3547860/bin/pntd.0002017.s002.docx
Authors & Co-Authors
Boubaker, Ghalia
Switzerland, Bern
University of Bern
Tunisia, Monastir
Université de Monastir
Macchiaroli, Natalia
Argentina, Buenos Aires
Universidad de Buenos Aires
Prada, Laura
Argentina, Buenos Aires
Universidad de Buenos Aires
Cucher, Marcela A.
Argentina, Buenos Aires
Universidad de Buenos Aires
Rosenzvit, Mara C.
Argentina, Buenos Aires
Universidad de Buenos Aires
Ziadinov, Iskender
Switzerland, Zurich
Universität Zürich
Deplazes, Peter
Switzerland, Zurich
Universität Zürich
Saarma, Urmas
Estonia, Tartu
Ökoloogia ja Maateaduste Instituut
Babba, Hamouda
Tunisia, Monastir
Université de Monastir
Gottstein, Bruno B.
Switzerland, Bern
University of Bern
Spiliotis, Markus
Switzerland, Bern
University of Bern
Statistics
Citations: 67
Authors: 11
Affiliations: 5
Identifiers
Doi:
10.1371/journal.pntd.0002017
ISSN:
19352727
e-ISSN:
19352735
Research Areas
Genetics And Genomics
Study Locations
Algeria
Tunisia