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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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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