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Microchip capillary electrophoresis-based genetic comparison of closely related cyathostomin nematode parasites of horses using randomly amplified polymorphic DNA polymerase chain reaction

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Abstract

The microchip-based capillary electrophoresis technology represents a valuable recent development for the analysis of complex DNA banding patterns. We have used this technology for the differentiation of the closely related cyathostomin species Cylicocyclus elongatus and C. insigne from the horse. We found that the Agilent 2100 bioanalyser in combination with the DNA 7500 Lab Chip were suited to perform a phylogenetic DNA fingerprinting analysis of the parasite species studied. The analysis of the electrophoretic data was optimised and it was possible to resolve a phylogenetic tree where all 12 individual worms of the two Cylicocyclus species studied were assigned to their species as determined by microscopic identification based on morphological traits. Thus, our data indicated that the procedure described here provides an additional powerful tool that can be employed for species delineation of closely related strains or species, such as the two taxa of Cylicocyclus investigated in the present study. Furthermore, by determining the second internal transcribed spacer region of three and nine individual worms for C. elongatus and C. insigne, respectively, low intraspecific variations of only up to 0.3% were demonstrated.

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Acknowledgements

The work reported here was supported by a grant from the Deutsche Forschungsgemeinschaft (Schn. 267/13–1). The authors wish to thank Dr Judith McAlister-Hermann and Dr William Blackhall for help in revising the manuscript.

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Authors and Affiliations

  1. Institute of Parasitology, Hannover School of Veterinary Medicine, Buenteweg 17, 30559, Hannover, Germany

    Janez Posedi, Thomas Schnieder, Johan Höglund, J. Ralph Lichtenfels & Georg von Samson-Himmelstjerna

  2. Institute of Microbiology and Parasitology, Veterinary Faculty of Ljubljana, University of Ljubljana, Ljubljana, Slovenia

    Janez Posedi

  3. Institute of Parasitology, Faculty of Veterinary Medicine, University of Leipzig, An den Tierkliniken 35, 04103, Leipzig, Germany

    Michaela Drögemüller

  4. Department of Parasitology (SWEPAR), National Veterinary Institute and Swedish University of Agriculture Sciences, Uppsala, Sweden

    Johan Höglund

  5. Animal Parasitic Diseases Laboratory, Beltsville Agricultural Research Center, Agricultural Research Service, U.S. Department of Agriculture, Beltsville, MD 20705–2350, USA

    J. Ralph Lichtenfels

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  1. Janez Posedi
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  2. Michaela Drögemüller
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  3. Thomas Schnieder
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  5. J. Ralph Lichtenfels
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  6. Georg von Samson-Himmelstjerna
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Correspondence to Georg von Samson-Himmelstjerna.

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Posedi, J., Drögemüller, M., Schnieder, T. et al. Microchip capillary electrophoresis-based genetic comparison of closely related cyathostomin nematode parasites of horses using randomly amplified polymorphic DNA polymerase chain reaction. Parasitol Res 92, 421–429 (2004). https://doi.org/10.1007/s00436-003-1067-3

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  • DOI: https://doi.org/10.1007/s00436-003-1067-3

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