PCR diagnosis of Fasciola hepatica in field-collected Lymnaea columella and Lymnaea viatrix snails

Fasciolosis, caused by the trematode Fasciola hepatica, is a zoonosis of economic importance in livestock that is emerging as a chronic disease in humans. The intermediate hosts are lymnaeid snails, in which diagnosis of infection is traditionally based on cercarial shedding, tissue sectioning and c...

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Autores principales: Cucher, M.A., Carnevale, S., Prepelitchi, L., Labbé, J.H., Wisnivesky-Colli, C.
Formato: JOUR
Materias:
PCR
Acceso en línea:http://hdl.handle.net/20.500.12110/paper_03044017_v137_n1-2_p74_Cucher
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spelling todo:paper_03044017_v137_n1-2_p74_Cucher2023-10-03T15:20:32Z PCR diagnosis of Fasciola hepatica in field-collected Lymnaea columella and Lymnaea viatrix snails Cucher, M.A. Carnevale, S. Prepelitchi, L. Labbé, J.H. Wisnivesky-Colli, C. Fasciola hepatica Lymnaea sp. PCR cytochrome c oxidase amplicon article controlled study cross reaction DNA sequence Fasciola hepatica infection rate Lymnaea lymnaea columella Lymnaea viatrix nonhuman nucleotide sequence polymerase chain reaction sensitivity and specificity snail Animals Argentina Base Sequence Cross Reactions Disease Reservoirs Disease Vectors DNA Primers Electron Transport Complex IV Fasciola hepatica Fascioliasis Lymnaea Polymerase Chain Reaction Sensitivity and Specificity Sequence Alignment Sequence Homology, Nucleic Acid Species Specificity Columella Fasciola hepatica Gastropoda Lymnaea Lymnaea columella Lymnaea viatrix Lymnaeidae Trematoda Viatrix Fasciolosis, caused by the trematode Fasciola hepatica, is a zoonosis of economic importance in livestock that is emerging as a chronic disease in humans. The intermediate hosts are lymnaeid snails, in which diagnosis of infection is traditionally based on cercarial shedding, tissue sectioning and crushing. We developed a PCR assay for the sensitive and specific detection of F. hepatica in field-collected Lymnaea sp. snails. A primer pair was designed to amplify a 405 bp fragment of the cytochrome c oxidase subunit 1 gene of F. hepatica. The PCR assay showed a limit of detection of 10 pg of genomic F. hepatica DNA. No cross-reactions were observed with samples from other related trematode species or from the snail hosts Lymnaea columella and Lymnaea viatrix. DNA sequencing of the amplicon showed 100% homology with F. hepatica, and 75-89% homology with other trematodes on regions that did not include the entire set of primers. Two samples from Argentina were analysed. For snails in sample 1 (n = 240), identified as L. columella, the infection rate was 17.5 and 51.3% by direct examination and PCR, respectively. For snails in sample 2 (n = 34), identified as L. viatrix, the infection rate was 2.9 and 61.8% by direct examination and PCR, respectively. Differences in infection rates between these diagnosis methods were significant for both samples. Our PCR technique showed to be effective for detecting specific F. hepatica infections of low intensity in the intermediate host, and hence it could be used to study the epidemiological situation in a given area, as well as to assess host suitability for the parasite. © 2005 Elsevier B.V. All rights reserved. Fil:Cucher, M.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Carnevale, S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Prepelitchi, L. 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_03044017_v137_n1-2_p74_Cucher
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-134
collection Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic Fasciola hepatica
Lymnaea sp.
PCR
cytochrome c oxidase
amplicon
article
controlled study
cross reaction
DNA sequence
Fasciola hepatica
infection rate
Lymnaea
lymnaea columella
Lymnaea viatrix
nonhuman
nucleotide sequence
polymerase chain reaction
sensitivity and specificity
snail
Animals
Argentina
Base Sequence
Cross Reactions
Disease Reservoirs
Disease Vectors
DNA Primers
Electron Transport Complex IV
Fasciola hepatica
Fascioliasis
Lymnaea
Polymerase Chain Reaction
Sensitivity and Specificity
Sequence Alignment
Sequence Homology, Nucleic Acid
Species Specificity
Columella
Fasciola hepatica
Gastropoda
Lymnaea
Lymnaea columella
Lymnaea viatrix
Lymnaeidae
Trematoda
Viatrix
spellingShingle Fasciola hepatica
Lymnaea sp.
PCR
cytochrome c oxidase
amplicon
article
controlled study
cross reaction
DNA sequence
Fasciola hepatica
infection rate
Lymnaea
lymnaea columella
Lymnaea viatrix
nonhuman
nucleotide sequence
polymerase chain reaction
sensitivity and specificity
snail
Animals
Argentina
Base Sequence
Cross Reactions
Disease Reservoirs
Disease Vectors
DNA Primers
Electron Transport Complex IV
Fasciola hepatica
Fascioliasis
Lymnaea
Polymerase Chain Reaction
Sensitivity and Specificity
Sequence Alignment
Sequence Homology, Nucleic Acid
Species Specificity
Columella
Fasciola hepatica
Gastropoda
Lymnaea
Lymnaea columella
Lymnaea viatrix
Lymnaeidae
Trematoda
Viatrix
Cucher, M.A.
Carnevale, S.
Prepelitchi, L.
Labbé, J.H.
Wisnivesky-Colli, C.
PCR diagnosis of Fasciola hepatica in field-collected Lymnaea columella and Lymnaea viatrix snails
topic_facet Fasciola hepatica
Lymnaea sp.
PCR
cytochrome c oxidase
amplicon
article
controlled study
cross reaction
DNA sequence
Fasciola hepatica
infection rate
Lymnaea
lymnaea columella
Lymnaea viatrix
nonhuman
nucleotide sequence
polymerase chain reaction
sensitivity and specificity
snail
Animals
Argentina
Base Sequence
Cross Reactions
Disease Reservoirs
Disease Vectors
DNA Primers
Electron Transport Complex IV
Fasciola hepatica
Fascioliasis
Lymnaea
Polymerase Chain Reaction
Sensitivity and Specificity
Sequence Alignment
Sequence Homology, Nucleic Acid
Species Specificity
Columella
Fasciola hepatica
Gastropoda
Lymnaea
Lymnaea columella
Lymnaea viatrix
Lymnaeidae
Trematoda
Viatrix
description Fasciolosis, caused by the trematode Fasciola hepatica, is a zoonosis of economic importance in livestock that is emerging as a chronic disease in humans. The intermediate hosts are lymnaeid snails, in which diagnosis of infection is traditionally based on cercarial shedding, tissue sectioning and crushing. We developed a PCR assay for the sensitive and specific detection of F. hepatica in field-collected Lymnaea sp. snails. A primer pair was designed to amplify a 405 bp fragment of the cytochrome c oxidase subunit 1 gene of F. hepatica. The PCR assay showed a limit of detection of 10 pg of genomic F. hepatica DNA. No cross-reactions were observed with samples from other related trematode species or from the snail hosts Lymnaea columella and Lymnaea viatrix. DNA sequencing of the amplicon showed 100% homology with F. hepatica, and 75-89% homology with other trematodes on regions that did not include the entire set of primers. Two samples from Argentina were analysed. For snails in sample 1 (n = 240), identified as L. columella, the infection rate was 17.5 and 51.3% by direct examination and PCR, respectively. For snails in sample 2 (n = 34), identified as L. viatrix, the infection rate was 2.9 and 61.8% by direct examination and PCR, respectively. Differences in infection rates between these diagnosis methods were significant for both samples. Our PCR technique showed to be effective for detecting specific F. hepatica infections of low intensity in the intermediate host, and hence it could be used to study the epidemiological situation in a given area, as well as to assess host suitability for the parasite. © 2005 Elsevier B.V. All rights reserved.
format JOUR
author Cucher, M.A.
Carnevale, S.
Prepelitchi, L.
Labbé, J.H.
Wisnivesky-Colli, C.
author_facet Cucher, M.A.
Carnevale, S.
Prepelitchi, L.
Labbé, J.H.
Wisnivesky-Colli, C.
author_sort Cucher, M.A.
title PCR diagnosis of Fasciola hepatica in field-collected Lymnaea columella and Lymnaea viatrix snails
title_short PCR diagnosis of Fasciola hepatica in field-collected Lymnaea columella and Lymnaea viatrix snails
title_full PCR diagnosis of Fasciola hepatica in field-collected Lymnaea columella and Lymnaea viatrix snails
title_fullStr PCR diagnosis of Fasciola hepatica in field-collected Lymnaea columella and Lymnaea viatrix snails
title_full_unstemmed PCR diagnosis of Fasciola hepatica in field-collected Lymnaea columella and Lymnaea viatrix snails
title_sort pcr diagnosis of fasciola hepatica in field-collected lymnaea columella and lymnaea viatrix snails
url http://hdl.handle.net/20.500.12110/paper_03044017_v137_n1-2_p74_Cucher
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