Diagnosis of fasciolosis in sheep using a loop-mediated isothermal amplification (LAMP) assay with faecal samples
| Autor: | Martínez Valladares, María, Rojo Vázquez, Francisco Antonio |
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| Rok vydání: | 2016 |
| Zdroj: | Digital.CSIC. Repositorio Institucional del CSIC instname |
| Popis: | Trabajo presentado al 12th European Multicolloquium of Parasitology (EMOP). Turku, (Finland), 20-24 julio 2016. In this study a loop-mediated isothermal amplification (LAMP) assay was developed with the aim to improve the simplicity and accuracy of Fasciola spp. diagnosis in faeces of sheep. LAMP assay is a very specific, efficient and rapid gene amplification procedure in which the reaction can run at a constant temperature. The primers used for the LAMP assay included an internal transcribed spacer 2 (ITS2) region; two sets of primers comprising two outer and two inner were selected. After the optimization of the assay, the limit of detection was 10 pg, the same as by a standard PCR with the outer primers. With the LAMP assay, we confirmed the infection in experimentally infected sheep from the first week, at the same time as using a standard PCR with the outer primers. In this study we also analysed the faeces of eight naturally infected sheep on day 0 and 30 after a treatment with triclabendazole. The infection by F. hepatica was confirmed in all animals before the treatment although on day 30 two sheep were diagnosed with fasciolosis using the LAMP assay. However, when these results were compared with the standard PCR using the outer primers, the results before treatment were the same but on day 30 we only confirmed the infection in one out of the two sheep. In conclusion, the LAMP assay could be a good alternative to conventional diagnostic methods to detect fasciolosis in faeces and also solves the drawbacks of the standard PCR. |
| Databáze: | OpenAIRE |
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