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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Autores principales:	Duffy, Tomás, Kleiman, Florencia, Pietrokovsky, Silvia Monica, Schijman, Alejandro Gabriel, Wisnivesky Colli, María Cristina
Publicado:	2009
Materias:	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 gene location genetic manipulation genetic polymorphism Lymnaea melting point nonhuman nucleotide sequence real time polymerase chain reaction species comparison species differentiation species identification Animals Phylogeny Polymerase Chain Reaction RNA, Ribosomal, 18S Sequence Analysis, DNA South America Diaphana Gastropoda
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_0001706X_v109_n1_p1_Duffy http://hdl.handle.net/20.500.12110/paper_0001706X_v109_n1_p1_Duffy
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_0001706X_v109_n1_p1_Duffy
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spelling	paper:paper_0001706X_v109_n1_p1_Duffy2023-06-08T14:21:07Z Real-time PCR strategy for rapid discrimination among main lymnaeid species from Argentina Duffy, Tomás Kleiman, Florencia Pietrokovsky, Silvia Monica Schijman, Alejandro Gabriel Wisnivesky Colli, María Cristina 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. 2009 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_0001706X_v109_n1_p1_Duffy 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
collection	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, Tomás Kleiman, Florencia Pietrokovsky, Silvia Monica Schijman, Alejandro Gabriel Wisnivesky Colli, María Cristina 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.
author	Duffy, Tomás Kleiman, Florencia Pietrokovsky, Silvia Monica Schijman, Alejandro Gabriel Wisnivesky Colli, María Cristina
author_facet	Duffy, Tomás Kleiman, Florencia Pietrokovsky, Silvia Monica Schijman, Alejandro Gabriel Wisnivesky Colli, María Cristina
author_sort	Duffy, Tomás
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
publishDate	2009
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_0001706X_v109_n1_p1_Duffy http://hdl.handle.net/20.500.12110/paper_0001706X_v109_n1_p1_Duffy
work_keys_str_mv	AT duffytomas realtimepcrstrategyforrapiddiscriminationamongmainlymnaeidspeciesfromargentina AT kleimanflorencia realtimepcrstrategyforrapiddiscriminationamongmainlymnaeidspeciesfromargentina AT pietrokovskysilviamonica realtimepcrstrategyforrapiddiscriminationamongmainlymnaeidspeciesfromargentina AT schijmanalejandrogabriel realtimepcrstrategyforrapiddiscriminationamongmainlymnaeidspeciesfromargentina AT wisniveskycollimariacristina realtimepcrstrategyforrapiddiscriminationamongmainlymnaeidspeciesfromargentina
_version_	1768542575148400640

Real-time PCR strategy for rapid discrimination among main lymnaeid species from Argentina

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