From complete mitochondrial DNA sequence of Fasciola hepatica
available in Genbank, specific primers were designed for a conserved and repetitive region of this trematode. A pair of primers was used for diagnosis of infected Lymnaea columella
by F. hepatica
during the pre-patent period simultaneously with another pair of primers which amplified the internal transcribed spacer (ITS) region of rDNA from L. columella
in a single Multiplex-PCR. The amplification generated a ladder band profile specific for F. hepatica
. This profile was observed in positive molluscs at different times of infection, including adult worms from the trematode. The Multiplex-PCR technique showed to be a fast and safe tool for fascioliasis diagnosis, enabling the detection of F. hepatica
miracidia in L. columella
during the pre-patent period and identification of transmission areas.