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.
