Development of a duplex PCR for the identification of Fasciola hepatica in lymnaeid snails
Fasciola hepatica is a parasitic trematode that causes fascioliasis, a disease that affects domestic livestock and humans. The complex life cycle of F. hepatica involves lymnaeid snails as intermediate hosts. Detection of F. hepatica in snails is a useful tool for the control of fascioliasis in live...
| Main Authors: | , , , |
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| Format: | info:ar-repo/semantics/artículo |
| Language: | Inglés |
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Ediciones INTA
2020
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| Subjects: | |
| Online Access: | http://ria.inta.gob.ar/contenido/ria-46-no-3-diciembre-2020 http://hdl.handle.net/20.500.12123/8522 |
| _version_ | 1855036051055181824 |
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| author | Mignaqui, Ana Clara Alvarez, Lucia Paula Soler, Paula Larroza, Marcela Patricia |
| author_browse | Alvarez, Lucia Paula Larroza, Marcela Patricia Mignaqui, Ana Clara Soler, Paula |
| author_facet | Mignaqui, Ana Clara Alvarez, Lucia Paula Soler, Paula Larroza, Marcela Patricia |
| author_sort | Mignaqui, Ana Clara |
| collection | INTA Digital |
| description | Fasciola hepatica is a parasitic trematode that causes fascioliasis, a disease that affects domestic livestock and humans. The complex life cycle of F. hepatica involves lymnaeid snails as intermediate hosts. Detection of F. hepatica in snails is a useful tool for the control of fascioliasis in livestock. Detection methods involve crushing the snails and microscopic observation, but have low sensitivity and are time-consuming. To overcome these disadvantages, researchers are developing molecular methods. In this work, we developed a duplex PCR that allows the detection of F. hepatica in snails as two single and bright bands: one corresponding to the parasite and one to the snail, the latter of which works as an internal control to detect PCR inhibitors. To avoid false-positive results, we also evaluated the method of disinfection of the material used for snail collection. The duplex PCR developed showed a sensitivity high enough to detect a single miracidium per snail, and significantly shortened the time required to analyze a large number of snails. |
| format | info:ar-repo/semantics/artículo |
| id | INTA8522 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2020 |
| publishDateRange | 2020 |
| publishDateSort | 2020 |
| publisher | Ediciones INTA |
| publisherStr | Ediciones INTA |
| record_format | dspace |
| spelling | INTA85222020-12-30T13:14:27Z Development of a duplex PCR for the identification of Fasciola hepatica in lymnaeid snails Mignaqui, Ana Clara Alvarez, Lucia Paula Soler, Paula Larroza, Marcela Patricia Fasciola hepatica Enfermedades de los Animales PCR Diagnóstico Animal Diseases Diagnosis Fasciolosis Caracoles Fasciola hepatica is a parasitic trematode that causes fascioliasis, a disease that affects domestic livestock and humans. The complex life cycle of F. hepatica involves lymnaeid snails as intermediate hosts. Detection of F. hepatica in snails is a useful tool for the control of fascioliasis in livestock. Detection methods involve crushing the snails and microscopic observation, but have low sensitivity and are time-consuming. To overcome these disadvantages, researchers are developing molecular methods. In this work, we developed a duplex PCR that allows the detection of F. hepatica in snails as two single and bright bands: one corresponding to the parasite and one to the snail, the latter of which works as an internal control to detect PCR inhibitors. To avoid false-positive results, we also evaluated the method of disinfection of the material used for snail collection. The duplex PCR developed showed a sensitivity high enough to detect a single miracidium per snail, and significantly shortened the time required to analyze a large number of snails. Fasciola hepatica es un parásito trematodo que causa fasciolosis, una enfermedad que afecta al ganado doméstico y al ser humano. El complejo ciclo de vida de F. hepatica involucra a los caracoles lymnaeidos como huéspedes intermediarios. La detección de F. hepatica en caracoles es una herramienta útil para el control de la fasciolosis en el ganado. Los métodos de detección implican el aplastamiento de los caracoles y la observación microscópica, pero tienen baja sensibilidad y consumen mucho tiempo. Para superar estas desventajas, se encuentran en desarrollo métodos de diagnóstico molecular. En este trabajo, se desarrolló una PCR dúplex que permite la detección de F. hepatica en caracoles como dos bandas simples y brillantes: una banda corresponde al parásito y otra al caracol, funcionando esta última como control interno para detectar inhibidores de la PCR. Para evitar resultados falsos positivos, también evaluamos el método de desinfección del material utilizado para la manipulación de caracoles. La PCR dúplex desarrollada mostró una sensibilidad lo suficientemente alta como para detectar un solo miracidio por caracol y acortó significativamente el tiempo de trabajo de análisis de una gran cantidad de caracoles. EEA Bariloche Fil: Mignaqui, Ana Clara. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Bariloche. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Instituto de Investigaciones Forestales y Agropecuarias Bariloche; Argentina Fil: Alvarez, Lucia Paula. Universidad Nacional del Comahue. Instituto Andino Patagónico de Tecnologías Biológicas y Geoambientales. Consejo Nacional de Investigaciones Científicas y Técnicas. 2 Instituto Andino Patagónico de Tecnologías Biológicas y Geoambientales; Argentina Fil: Soler, Paula. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Bariloche. Instituto de Investigaciones Forestales y Agropecuarias Bariloche. Grupo Sanidad Animal; Argentina Fil: Larroza, Marcela Patricia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Bariloche. Instituto de Investigaciones Forestales y Agropecuarias Bariloche. Grupo Sanidad Animal; Argentina 2020-12-30T12:57:22Z 2020-12-30T12:57:22Z 2020-12 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://ria.inta.gob.ar/contenido/ria-46-no-3-diciembre-2020 http://hdl.handle.net/20.500.12123/8522 0325-8718 1669-2314 eng info:eu-repograntAgreement/INTA/PATNOR-1281101/AR./Aportes para el desarrollo de cordillera y precordillera. info:eu-repograntAgreement/INTA/PATNOR-1281103/AR./Contribución al desarrollo regional del centro y norte neuquino. info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Ediciones INTA RIA 46 (3) : 347-351 (Diciembre 2020) |
| spellingShingle | Fasciola hepatica Enfermedades de los Animales PCR Diagnóstico Animal Diseases Diagnosis Fasciolosis Caracoles Mignaqui, Ana Clara Alvarez, Lucia Paula Soler, Paula Larroza, Marcela Patricia Development of a duplex PCR for the identification of Fasciola hepatica in lymnaeid snails |
| title | Development of a duplex PCR for the identification of Fasciola hepatica in lymnaeid snails |
| title_full | Development of a duplex PCR for the identification of Fasciola hepatica in lymnaeid snails |
| title_fullStr | Development of a duplex PCR for the identification of Fasciola hepatica in lymnaeid snails |
| title_full_unstemmed | Development of a duplex PCR for the identification of Fasciola hepatica in lymnaeid snails |
| title_short | Development of a duplex PCR for the identification of Fasciola hepatica in lymnaeid snails |
| title_sort | development of a duplex pcr for the identification of fasciola hepatica in lymnaeid snails |
| topic | Fasciola hepatica Enfermedades de los Animales PCR Diagnóstico Animal Diseases Diagnosis Fasciolosis Caracoles |
| url | http://ria.inta.gob.ar/contenido/ria-46-no-3-diciembre-2020 http://hdl.handle.net/20.500.12123/8522 |
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