Electrophoretic analysis of PCR products obtained with DNA of the adult trematodes: Lane M, DNA ladder (bp); lane 1, O. felineus DNA; lane 2, M. bilis DNA; lane 3, negative control. 

ITS2 fragments alignment indicating nucleotide positions used for primers design. 

Electrophoretic analysis of PCR products of O. felineus and M. bilis specimens from different definitive hosts: Lane M, DNA ladder (bp); lane 1, O. felineus (ides); lane 2, O. felineus (golden hamster); lane 3, O. felineus (red fox); lane 5, M. bilis (ides); lane 6, M. bilis (golden hamster); lane 7, M. bilis (red fox); lane 4, 8, negative control. 

Results of cross-reactivity test of liver flukes DNA and species-specific primers: Lane M, DNA ladder (bp); lane 1, O. felineus DNA and OF-spec-F/R primers; lane 2, O. felineus DNA and MB-spec-F/R primers; lane 4, M. bilis DNA and MB-spec-F/R primers; lane 5, M. bilis DNA and OF-spec-F/R primers; lane 3, 6, negative control. 

Results of cross-reactivity test of the intermediate host and different metacercariae DNA and species-specific primers: Lane M, DNA ladder (bp); lane 1, O. felineus; lane 2, M. bilis; lane 3, M. xanthosomus; lane 4, H. dubinini; lane 5, P. cuticola; lane 6, L. idus; lane 7, negative control. 

Results of cross-reactivity test of the intermediate host and different helminthes DNA and species-specific primers: Lane M, DNA ladder (bp); lane 1, O. felineus; lane 2, M. bilis; lane 3, T. spiralis; lane 4, T. canis; lane 5, T. hydatigena; lane 6, red fox DNA; lane 7, negative control. 

Results of the sensitivity test of species-specific primers with (A) O. felineus DNA and (B) M. bilis DNA: Lane M, DNA ladder (bp); lane 1, 20 ng; lane 2, 10 ng; lane 3, 8 ng; lane 4, 6 ng; lane 5, 4 ng; lane 6, 2 ng; lane 7, 1 ng; lane 8, negative control.