Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati
Abstract 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 t...
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Cambridge University Press (CUP)
2021
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Online Access: | http://dx.doi.org/10.1017/s0031182021000342 https://www.cambridge.org/core/services/aop-cambridge-core/content/view/S0031182021000342 |
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crcambridgeupr:10.1017/s0031182021000342 2024-09-15T18:01:22+00:00 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 Analía Butti, Marcos Javier Trangoni, Marcos David Grune Löffler, Sylvia Pérez, Verónica Mirtha Periago, María Victoria 2021 http://dx.doi.org/10.1017/s0031182021000342 https://www.cambridge.org/core/services/aop-cambridge-core/content/view/S0031182021000342 en eng Cambridge University Press (CUP) https://www.cambridge.org/core/terms Parasitology volume 148, issue 7, page 819-826 ISSN 0031-1820 1469-8161 journal-article 2021 crcambridgeupr https://doi.org/10.1017/s0031182021000342 2024-07-31T04:04:33Z Abstract 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. Article in Journal/Newspaper Canis lupus Cambridge University Press Parasitology 148 7 819 826 |
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Cambridge University Press |
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crcambridgeupr |
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English |
description |
Abstract 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. |
format |
Article in Journal/Newspaper |
author |
Avila, Héctor Gabriel Risso, Marikena Guadalupe Ruybal, Paula Repetto, Silvia Analía Butti, Marcos Javier Trangoni, Marcos David Grune Löffler, Sylvia Pérez, Verónica Mirtha Periago, María Victoria |
spellingShingle |
Avila, Héctor Gabriel Risso, Marikena Guadalupe Ruybal, Paula Repetto, Silvia Analía Butti, Marcos Javier Trangoni, Marcos David Grune Löffler, Sylvia Pérez, Verónica Mirtha Periago, María Victoria Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati |
author_facet |
Avila, Héctor Gabriel Risso, Marikena Guadalupe Ruybal, Paula Repetto, Silvia Analía Butti, Marcos Javier Trangoni, Marcos David Grune Löffler, Sylvia Pérez, Verónica Mirtha Periago, María 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 (CUP) |
publishDate |
2021 |
url |
http://dx.doi.org/10.1017/s0031182021000342 https://www.cambridge.org/core/services/aop-cambridge-core/content/view/S0031182021000342 |
genre |
Canis lupus |
genre_facet |
Canis lupus |
op_source |
Parasitology volume 148, issue 7, page 819-826 ISSN 0031-1820 1469-8161 |
op_rights |
https://www.cambridge.org/core/terms |
op_doi |
https://doi.org/10.1017/s0031182021000342 |
container_title |
Parasitology |
container_volume |
148 |
container_issue |
7 |
container_start_page |
819 |
op_container_end_page |
826 |
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1810438515843923968 |