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

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Abstract

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.

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Acknowledgements

For the provision of trematodes and affected livers, our thanks to Dr. B. Specht, Hohenfinow (muskrat), Institute for Veterinary Pathology, Freie Universität Berlin (domestic cat), Institute for Food, Drugs and Animal Epidemics, Berlin (red fox), Prof. Iljinskich, Medical University, Tomsk, Russia (human), Prof. Beer, Academy of Science, Moscow, Russia (moor buzzard). The experimental infection of animals was undertaken with the permission of the Regional Office for Industrial Safety, Health Protection and Technical Safety, Berlin (Reg. no: L0016/00 and 0049/01). The results are part of an ongoing dissertation at the Institute for Parasitology and International Animal Health (Freie Universität Berlin).

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Authors and Affiliations

  1. Institute for Parasitology and International Animal Health, Freie Universität Berlin, Berlin, Germany

    Andreas Pauly, Rolf Schuster & Stephan Steuber

  2. Königsweg 67, 14163, Berlin, Germany

    Andreas Pauly

  3. Central Veterinary Research Laboratory, P.O. B. 597, Dubai, United Arab Emirates

    Rolf Schuster

  4. Federal Office for Consumer Protection and Food Safety, Diedersdorfer Weg 1, 12277, Berlin, Germany

    Stephan Steuber

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  1. Andreas Pauly
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  2. Rolf Schuster
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  3. Stephan Steuber
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Correspondence to Andreas Pauly.

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Pauly, A., Schuster, R. & Steuber, S. Molecular characterization and differentiation of opisthorchiid trematodes of the species Opisthorchis felineus (Rivolta, 1884) and Metorchis bilis (Braun, 1790) using polymerase chain reaction. Parasitol Res 90, 409–414 (2003). https://doi.org/10.1007/s00436-003-0851-4

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  • DOI: https://doi.org/10.1007/s00436-003-0851-4

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