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

Fasciola hepatica is a parasitic trematode that causes fasciolosis, 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 fasciolosis in livestock. Detection methods involve crushing of 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 working time to analyze a large number of snails.