Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati
Toxocariasis is a zoonotic disease caused mainly by Toxocara canis and Toxocara cati and diagnosis in dogs and cats is an important tool for its control. For this reason, a new coprological loop-mediated isothermal amplification (LAMP) assay was developed for the simultaneous detection of these spec...
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ftconicet:oai:ri.conicet.gov.ar:11336/151870 2023-10-09T21:50:39+02:00 Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati Avila, Héctor Gabriel Risso, Marikena Guadalupe Ruybal, Paula Repetto, Silvia Butti, Marcos Javier Trangoni, Marcos David Grune Loffler, Sylvia Perez, Veronica Mirtha Periago, Maria Victoria application/pdf http://hdl.handle.net/11336/151870 eng eng Cambridge University Press info:eu-repo/semantics/altIdentifier/url/https://www.cambridge.org/core/product/identifier/S0031182021000342/type/journal_article info:eu-repo/semantics/altIdentifier/doi/10.1017/S0031182021000342 http://hdl.handle.net/11336/151870 Avila, Héctor Gabriel; Risso, Marikena Guadalupe; Ruybal, Paula; Repetto, Silvia; Butti, Marcos Javier; et al.; Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati; Cambridge University Press; Parasitology; 148; 7; 6-2021; 819-826 0031-1820 1469-8161 CONICET Digital CONICET info:eu-repo/semantics/openAccess https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ COPRO-DIAGNOSIS LOOP-MEDIATED ISOTHERMAL AMPLIFICATION LOW-COST TOXOCARA CANIS TOXOCARA CATI TOXOCARIASIS https://purl.org/becyt/ford/1.6 https://purl.org/becyt/ford/1 info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion ftconicet https://doi.org/10.1017/S0031182021000342 2023-09-24T18:51:02Z Toxocariasis is a zoonotic disease caused mainly by Toxocara canis and Toxocara cati and diagnosis in dogs and cats is an important tool for its control. For this reason, a new coprological loop-mediated isothermal amplification (LAMP) assay was developed for the simultaneous detection of these species. The primer set was designed on a region of the mitochondrial cox-1 gene. Amplification conditions were evaluated using a temperature gradient (52°C to 68°C), different incubation times (15-120 min), and different concentrations of malachite green dye (0.004-0.4% w/v). The analytical sensitivity was evaluated with serial dilutions of genomic DNA from T. canis and T. cati adult worms, and with serial dilutions of DNA extracted from feces using a low-cost in-house method. The specificity was evaluated using genomic DNA from Canis lupus familiaris, Felis catus, Escherichia coli, Toxascaris leonina, Ancylostoma caninum, Echinococcus granulosus sensu stricto and Taenia hydatigena. The LAMP assay applied to environmental fecal samples from an endemic area showed an analytical sensitivity of 10-100 fg of genomic DNA and 10-5 serial dilutions of DNA extracted from feces using the low-cost in-house method; with a specificity of 100%. Additionally, the total development of the assay was carried out in a basic laboratory and per-reaction reagent cost decreased by ~80%. This new, low-cost tool can help identify the most common agents of toxocariasis in endemic areas in order to manage prevention strategies without having to rely on a laboratory with sophisticated equipment. Fil: Avila, Héctor Gabriel. Universidad Católica de Cuyo. Facultad de Ciencias Químicas y Tecnológicas. Laboratorio Provincial de Zoonosis Provincial; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis; Argentina Fil: Risso, Marikena Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en ... Article in Journal/Newspaper Canis lupus CONICET Digital (Consejo Nacional de Investigaciones Científicas y Técnicas) Argentina Parasitology 148 7 819 826 |
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CONICET Digital (Consejo Nacional de Investigaciones Científicas y Técnicas) |
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ftconicet |
language |
English |
topic |
COPRO-DIAGNOSIS LOOP-MEDIATED ISOTHERMAL AMPLIFICATION LOW-COST TOXOCARA CANIS TOXOCARA CATI TOXOCARIASIS https://purl.org/becyt/ford/1.6 https://purl.org/becyt/ford/1 |
spellingShingle |
COPRO-DIAGNOSIS LOOP-MEDIATED ISOTHERMAL AMPLIFICATION LOW-COST TOXOCARA CANIS TOXOCARA CATI TOXOCARIASIS https://purl.org/becyt/ford/1.6 https://purl.org/becyt/ford/1 Avila, Héctor Gabriel Risso, Marikena Guadalupe Ruybal, Paula Repetto, Silvia Butti, Marcos Javier Trangoni, Marcos David Grune Loffler, Sylvia Perez, Veronica Mirtha Periago, Maria Victoria Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati |
topic_facet |
COPRO-DIAGNOSIS LOOP-MEDIATED ISOTHERMAL AMPLIFICATION LOW-COST TOXOCARA CANIS TOXOCARA CATI TOXOCARIASIS https://purl.org/becyt/ford/1.6 https://purl.org/becyt/ford/1 |
description |
Toxocariasis is a zoonotic disease caused mainly by Toxocara canis and Toxocara cati and diagnosis in dogs and cats is an important tool for its control. For this reason, a new coprological loop-mediated isothermal amplification (LAMP) assay was developed for the simultaneous detection of these species. The primer set was designed on a region of the mitochondrial cox-1 gene. Amplification conditions were evaluated using a temperature gradient (52°C to 68°C), different incubation times (15-120 min), and different concentrations of malachite green dye (0.004-0.4% w/v). The analytical sensitivity was evaluated with serial dilutions of genomic DNA from T. canis and T. cati adult worms, and with serial dilutions of DNA extracted from feces using a low-cost in-house method. The specificity was evaluated using genomic DNA from Canis lupus familiaris, Felis catus, Escherichia coli, Toxascaris leonina, Ancylostoma caninum, Echinococcus granulosus sensu stricto and Taenia hydatigena. The LAMP assay applied to environmental fecal samples from an endemic area showed an analytical sensitivity of 10-100 fg of genomic DNA and 10-5 serial dilutions of DNA extracted from feces using the low-cost in-house method; with a specificity of 100%. Additionally, the total development of the assay was carried out in a basic laboratory and per-reaction reagent cost decreased by ~80%. This new, low-cost tool can help identify the most common agents of toxocariasis in endemic areas in order to manage prevention strategies without having to rely on a laboratory with sophisticated equipment. Fil: Avila, Héctor Gabriel. Universidad Católica de Cuyo. Facultad de Ciencias Químicas y Tecnológicas. Laboratorio Provincial de Zoonosis Provincial; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis; Argentina Fil: Risso, Marikena Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones en ... |
format |
Article in Journal/Newspaper |
author |
Avila, Héctor Gabriel Risso, Marikena Guadalupe Ruybal, Paula Repetto, Silvia Butti, Marcos Javier Trangoni, Marcos David Grune Loffler, Sylvia Perez, Veronica Mirtha Periago, Maria Victoria |
author_facet |
Avila, Héctor Gabriel Risso, Marikena Guadalupe Ruybal, Paula Repetto, Silvia Butti, Marcos Javier Trangoni, Marcos David Grune Loffler, Sylvia Perez, Veronica Mirtha Periago, Maria Victoria |
author_sort |
Avila, Héctor Gabriel |
title |
Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati |
title_short |
Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati |
title_full |
Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati |
title_fullStr |
Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati |
title_full_unstemmed |
Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati |
title_sort |
development of a low-cost copro-lamp assay for simultaneous copro-detection of toxocara canis and toxocara cati |
publisher |
Cambridge University Press |
url |
http://hdl.handle.net/11336/151870 |
geographic |
Argentina |
geographic_facet |
Argentina |
genre |
Canis lupus |
genre_facet |
Canis lupus |
op_relation |
info:eu-repo/semantics/altIdentifier/url/https://www.cambridge.org/core/product/identifier/S0031182021000342/type/journal_article info:eu-repo/semantics/altIdentifier/doi/10.1017/S0031182021000342 http://hdl.handle.net/11336/151870 Avila, Héctor Gabriel; Risso, Marikena Guadalupe; Ruybal, Paula; Repetto, Silvia; Butti, Marcos Javier; et al.; Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati; Cambridge University Press; Parasitology; 148; 7; 6-2021; 819-826 0031-1820 1469-8161 CONICET Digital CONICET |
op_rights |
info:eu-repo/semantics/openAccess https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ |
op_doi |
https://doi.org/10.1017/S0031182021000342 |
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Parasitology |
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148 |
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7 |
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819 |
op_container_end_page |
826 |
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1779313716889649152 |