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Artículo

Development of a low-cost copro-LAMP assay for simultaneous copro-detection of Toxocara canis and Toxocara cati

Avila, Héctor GabrielIcon ; Risso, Marikena GuadalupeIcon ; Ruybal, PaulaIcon ; Repetto, Silvia; Butti, Marcos Javier; Trangoni, Marcos David; Grune Loffler, SylviaIcon ; Perez, Veronica Mirtha; Periago, Maria VictoriaIcon
Fecha de publicación: 06/2021
Editorial: Cambridge University Press
Revista: Parasitology
ISSN: 0031-1820
e-ISSN: 1469-8161
Idioma: Inglés
Tipo de recurso: Artículo publicado
Clasificación temática:
Bioquímica y Biología Molecular

Resumen

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.
Palabras clave: COPRO-DIAGNOSIS , LOOP-MEDIATED ISOTHERMAL AMPLIFICATION , LOW-COST , TOXOCARA CANIS , TOXOCARA CATI , TOXOCARIASIS
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info:eu-repo/semantics/openAccess Excepto donde se diga explícitamente, este item se publica bajo la siguiente descripción: Creative Commons Attribution-NonCommercial-ShareAlike 2.5 Unported (CC BY-NC-SA 2.5)
Identificadores
URI: http://hdl.handle.net/11336/151870
URL: https://www.cambridge.org/core/product/identifier/S0031182021000342/type/journal
DOI: https://doi.org/10.1017/S0031182021000342
Colecciones
Articulos(CCT - SAN JUAN)
Articulos de CENTRO CIENTIFICO TECNOLOGICO CONICET - SAN JUAN
Articulos(IMPAM)
Articulos de INSTITUTO DE INVESTIGACIONES EN MICROBIOLOGIA Y PARASITOLOGIA MEDICA
Articulos(SEDE CENTRAL)
Articulos de SEDE CENTRAL
Citación
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
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