Real-time PCR strategy for rapid discrimination among main lymnaeid species from Argentina
Snails of the Family Lymnaeidae act as an intermediate hosts of Fasciola hepatica worldwide. The taxonomy of lymnaeid species is relevant for epidemiological studies and molecular strategies are increasingly used for that purpose. This work presents the first report of a real-time PCR approach used...
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todo:paper_0001706X_v109_n1_p1_Duffy2023-10-03T13:51:35Z Real-time PCR strategy for rapid discrimination among main lymnaeid species from Argentina Duffy, T. Kleiman, F. Pietrokovsky, S. Issia, L. Schijman, A.G. Wisnivesky-Colli, C. 18S rDNA Fasciolosis Galba truncatula Lymnaea diaphana Lymnaea viatrix Lymnaeidae Pseudosuccinea columella Real-time PCR ribosome RNA endemic species flatworm identification method intermediate host polymerase chain reaction polymorphism real time snail taxonomy amplicon Argentina article control strategy endemic disease Fasciola hepatica Galba truncatula gene location genetic manipulation genetic polymorphism intermediate host Lymnaea Lymnaea diaphana Lymnaea viatrix melting point nonhuman nucleotide sequence Pseudosuccinea columella real time polymerase chain reaction species comparison species differentiation species identification taxonomy Animals Argentina Lymnaea Phylogeny Polymerase Chain Reaction RNA, Ribosomal, 18S Sequence Analysis, DNA Argentina South America Diaphana Fasciola hepatica Galba truncatula Gastropoda Lymnaea Lymnaea viatrix Lymnaeidae Pseudosuccinea columella Snails of the Family Lymnaeidae act as an intermediate hosts of Fasciola hepatica worldwide. The taxonomy of lymnaeid species is relevant for epidemiological studies and molecular strategies are increasingly used for that purpose. This work presents the first report of a real-time PCR approach used to identify the most important lymnaeid species in the Southern Cone of South America. Species discrimination is based on the sequence polymorphism located within the helix E10-1 of the variable region V2 of the 18S rRNA genes, which yields amplicons with clearly different melting temperatures. This procedure minimises the risk of carry-over contamination because it does not require post-PCR manipulations, and the whole protocol can be completed in less than 4 h with a single snail foot as starting material. This method was successfully carried out in a blind study that included a panel of 20 Galba truncatula, 5 Lymnaea viatrix, 5 Lymnaea diaphana and 5 Pseudosuccinea columella specimens from different endemic areas for fasciolosis. This molecular approach constitutes a key laboratory tool complementing ecological studies that ultimately will promote more efficient control strategies. © 2008 Elsevier B.V. All rights reserved. Fil:Duffy, T. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Kleiman, F. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Pietrokovsky, S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Schijman, A.G. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Wisnivesky-Colli, C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_0001706X_v109_n1_p1_Duffy |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
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Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
18S rDNA Fasciolosis Galba truncatula Lymnaea diaphana Lymnaea viatrix Lymnaeidae Pseudosuccinea columella Real-time PCR ribosome RNA endemic species flatworm identification method intermediate host polymerase chain reaction polymorphism real time snail taxonomy amplicon Argentina article control strategy endemic disease Fasciola hepatica Galba truncatula gene location genetic manipulation genetic polymorphism intermediate host Lymnaea Lymnaea diaphana Lymnaea viatrix melting point nonhuman nucleotide sequence Pseudosuccinea columella real time polymerase chain reaction species comparison species differentiation species identification taxonomy Animals Argentina Lymnaea Phylogeny Polymerase Chain Reaction RNA, Ribosomal, 18S Sequence Analysis, DNA Argentina South America Diaphana Fasciola hepatica Galba truncatula Gastropoda Lymnaea Lymnaea viatrix Lymnaeidae Pseudosuccinea columella |
| spellingShingle |
18S rDNA Fasciolosis Galba truncatula Lymnaea diaphana Lymnaea viatrix Lymnaeidae Pseudosuccinea columella Real-time PCR ribosome RNA endemic species flatworm identification method intermediate host polymerase chain reaction polymorphism real time snail taxonomy amplicon Argentina article control strategy endemic disease Fasciola hepatica Galba truncatula gene location genetic manipulation genetic polymorphism intermediate host Lymnaea Lymnaea diaphana Lymnaea viatrix melting point nonhuman nucleotide sequence Pseudosuccinea columella real time polymerase chain reaction species comparison species differentiation species identification taxonomy Animals Argentina Lymnaea Phylogeny Polymerase Chain Reaction RNA, Ribosomal, 18S Sequence Analysis, DNA Argentina South America Diaphana Fasciola hepatica Galba truncatula Gastropoda Lymnaea Lymnaea viatrix Lymnaeidae Pseudosuccinea columella Duffy, T. Kleiman, F. Pietrokovsky, S. Issia, L. Schijman, A.G. Wisnivesky-Colli, C. Real-time PCR strategy for rapid discrimination among main lymnaeid species from Argentina |
| topic_facet |
18S rDNA Fasciolosis Galba truncatula Lymnaea diaphana Lymnaea viatrix Lymnaeidae Pseudosuccinea columella Real-time PCR ribosome RNA endemic species flatworm identification method intermediate host polymerase chain reaction polymorphism real time snail taxonomy amplicon Argentina article control strategy endemic disease Fasciola hepatica Galba truncatula gene location genetic manipulation genetic polymorphism intermediate host Lymnaea Lymnaea diaphana Lymnaea viatrix melting point nonhuman nucleotide sequence Pseudosuccinea columella real time polymerase chain reaction species comparison species differentiation species identification taxonomy Animals Argentina Lymnaea Phylogeny Polymerase Chain Reaction RNA, Ribosomal, 18S Sequence Analysis, DNA Argentina South America Diaphana Fasciola hepatica Galba truncatula Gastropoda Lymnaea Lymnaea viatrix Lymnaeidae Pseudosuccinea columella |
| description |
Snails of the Family Lymnaeidae act as an intermediate hosts of Fasciola hepatica worldwide. The taxonomy of lymnaeid species is relevant for epidemiological studies and molecular strategies are increasingly used for that purpose. This work presents the first report of a real-time PCR approach used to identify the most important lymnaeid species in the Southern Cone of South America. Species discrimination is based on the sequence polymorphism located within the helix E10-1 of the variable region V2 of the 18S rRNA genes, which yields amplicons with clearly different melting temperatures. This procedure minimises the risk of carry-over contamination because it does not require post-PCR manipulations, and the whole protocol can be completed in less than 4 h with a single snail foot as starting material. This method was successfully carried out in a blind study that included a panel of 20 Galba truncatula, 5 Lymnaea viatrix, 5 Lymnaea diaphana and 5 Pseudosuccinea columella specimens from different endemic areas for fasciolosis. This molecular approach constitutes a key laboratory tool complementing ecological studies that ultimately will promote more efficient control strategies. © 2008 Elsevier B.V. All rights reserved. |
| format |
JOUR |
| author |
Duffy, T. Kleiman, F. Pietrokovsky, S. Issia, L. Schijman, A.G. Wisnivesky-Colli, C. |
| author_facet |
Duffy, T. Kleiman, F. Pietrokovsky, S. Issia, L. Schijman, A.G. Wisnivesky-Colli, C. |
| author_sort |
Duffy, T. |
| title |
Real-time PCR strategy for rapid discrimination among main lymnaeid species from Argentina |
| title_short |
Real-time PCR strategy for rapid discrimination among main lymnaeid species from Argentina |
| title_full |
Real-time PCR strategy for rapid discrimination among main lymnaeid species from Argentina |
| title_fullStr |
Real-time PCR strategy for rapid discrimination among main lymnaeid species from Argentina |
| title_full_unstemmed |
Real-time PCR strategy for rapid discrimination among main lymnaeid species from Argentina |
| title_sort |
real-time pcr strategy for rapid discrimination among main lymnaeid species from argentina |
| url |
http://hdl.handle.net/20.500.12110/paper_0001706X_v109_n1_p1_Duffy |
| work_keys_str_mv |
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