Discrimination of the ITS1 types of Fasciola spp. based on a PCR–RFLP method (original) (raw)

Abstract

Molecular characterization is important for discriminating Fasciola specimens having the deoxyribonucleic acid (DNA) sequences of Fasciola hepatica, Fasciola gigantica, and both Fasciola species, since three Fasciola forms coexist in Asian countries. We have developed a restriction fragment length polymorphism of amplified DNA (PCR–RFLP) of the nuclear ribosomal internal transcribed spacer 1 (ITS1) region in Fasciola species. The band patterns of the fragments digested with a restriction enzyme, Rsa I, were accurately distinguished among the three forms of Fasciola. Amplicons with the sequences of F. hepatica and F. gigantica were divided into fragments of about 360, 100, and 60 bp, and 360, 170, and 60 bp, respectively, and amplicons with the sequences of both Fasciola species yielded fragments of 360, 170, 100, and 60 bp. The results of PCR–RFLP completely coincided with those of sequence analysis, and thus PCR–RFLP is a useful technique for determining the ITS1 type in Fasciola species.

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Acknowledgements

This study was supported in part by a grant-in-aid for Scientific Research (B) (no. 18405035) from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

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Authors and Affiliations

1. Laboratory of Veterinary Parasitology, Faculty of Agriculture, Iwate University, Ueda 3-18-8, Morioka, 020-8550, Japan
   Madoka Ichikawa & Tadashi Itagaki

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1. Madoka Ichikawa
2. Tadashi Itagaki

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Correspondence toTadashi Itagaki.

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Ichikawa, M., Itagaki, T. Discrimination of the ITS1 types of Fasciola spp. based on a PCR–RFLP method.Parasitol Res 106, 757–761 (2010). https://doi.org/10.1007/s00436-010-1724-2

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• Received: 21 October 2009
• Accepted: 04 January 2010
• Published: 28 January 2010
• Issue date: February 2010
• DOI: https://doi.org/10.1007/s00436-010-1724-2

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