Molecular characterization and differentiation of opisthorchiid trematodes of the species Opisthorchis felineus (Rivolta, 1884) and Metorchis bilis (Braun, 1790) using polymerase chain reaction

Adult specimens of the opisthorchiid liver fluke species Opisthorchis felineus and Metorchis bilis could be identified for the first time by molecular biological methods using species specific primers (OF and MB primers) in the polymerase chain reaction (PCR). The OF or MB primers were based on a part of the mitochondrial cytochrome c oxidase I gene. A specific product of approximately 200 bp could be amplified for O. felineus by means of the specific O. felineus primers. By contrast, the amplification of M. bilis DNA with MB primers produced a fragment of approximately 110 bp. A specificity of 100% could be demonstrated for both primer pairs. The sensitivities of the PCRs were 10 pg for the O. felineus DNA and 100 fg for the M. bilis DNA.