Multiple Alignment of the 28 large subunit of ribosomal RNA from 5 fishborne trematodes. Alignment of (28S rRNA) genes of Opisthorchis viverrini (HM004188), Clonorchis sinensis (JF823989), Haplorchis taichui (HM004187), H. pumilio (HM004191), and Stellantchasmus falcatus (KF241630) available in GenBank were selected to find a suitable region for discrimination these five species. The solid arrows indicate the position of OVPyro28S_F (forward primer) and biotinylated OVPyro28S_R (reverse primer) for template amplification. A dash arrow indicates OVPyro28S_S (sequencing primer), and a rectangular box shows the position of the 47 nucleotides as the target region used for species level identification. Dots indicate identical nucleotides between lines. Dash indicates gap between nucleotides.