A multiplex PCR for the simultaneous detection and genotyping of the Echinococcus granulosus complex.
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. canade...
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ftdoajarticles:oai:doaj.org/article:140181023b444a45acf161881c792116 2023-05-15T15:13:19+02:00 A multiplex PCR for the simultaneous detection and genotyping of the Echinococcus granulosus complex. Ghalia Boubaker Natalia Macchiaroli Laura Prada Marcela A Cucher Mara C Rosenzvit Iskender Ziadinov Peter Deplazes Urmas Saarma Hamouda Babba Bruno Gottstein Markus Spiliotis 2013-01-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0002017 https://doaj.org/article/140181023b444a45acf161881c792116 EN eng Public Library of Science (PLoS) https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23350011/pdf/?tool=EBI https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0002017 https://doaj.org/article/140181023b444a45acf161881c792116 PLoS Neglected Tropical Diseases, Vol 7, Iss 1, p e2017 (2013) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2013 ftdoajarticles https://doi.org/10.1371/journal.pntd.0002017 2022-12-31T05:47:44Z 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. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Argentina PLoS Neglected Tropical Diseases 7 1 e2017 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
spellingShingle |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Ghalia Boubaker Natalia Macchiaroli Laura Prada Marcela A Cucher Mara C Rosenzvit Iskender Ziadinov Peter Deplazes Urmas Saarma Hamouda Babba Bruno Gottstein Markus Spiliotis A multiplex PCR for the simultaneous detection and genotyping of the Echinococcus granulosus complex. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
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. |
format |
Article in Journal/Newspaper |
author |
Ghalia Boubaker Natalia Macchiaroli Laura Prada Marcela A Cucher Mara C Rosenzvit Iskender Ziadinov Peter Deplazes Urmas Saarma Hamouda Babba Bruno Gottstein Markus Spiliotis |
author_facet |
Ghalia Boubaker Natalia Macchiaroli Laura Prada Marcela A Cucher Mara C Rosenzvit Iskender Ziadinov Peter Deplazes Urmas Saarma Hamouda Babba Bruno Gottstein Markus Spiliotis |
author_sort |
Ghalia Boubaker |
title |
A multiplex PCR for the simultaneous detection and genotyping of the Echinococcus granulosus complex. |
title_short |
A multiplex PCR for the simultaneous detection and genotyping of the Echinococcus granulosus complex. |
title_full |
A multiplex PCR for the simultaneous detection and genotyping of the Echinococcus granulosus complex. |
title_fullStr |
A multiplex PCR for the simultaneous detection and genotyping of the Echinococcus granulosus complex. |
title_full_unstemmed |
A multiplex PCR for the simultaneous detection and genotyping of the Echinococcus granulosus complex. |
title_sort |
multiplex pcr for the simultaneous detection and genotyping of the echinococcus granulosus complex. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2013 |
url |
https://doi.org/10.1371/journal.pntd.0002017 https://doaj.org/article/140181023b444a45acf161881c792116 |
geographic |
Arctic Argentina |
geographic_facet |
Arctic Argentina |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 7, Iss 1, p e2017 (2013) |
op_relation |
https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23350011/pdf/?tool=EBI https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0002017 https://doaj.org/article/140181023b444a45acf161881c792116 |
op_doi |
https://doi.org/10.1371/journal.pntd.0002017 |
container_title |
PLoS Neglected Tropical Diseases |
container_volume |
7 |
container_issue |
1 |
container_start_page |
e2017 |
_version_ |
1766343885223624704 |