Development of a duplex PCR for the identification of Fasciola hepatica in lymnaeid snails

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Instituto Nacional de Tecnología Agropecuaria

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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.

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