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A fluorescence-based polymerase chain reaction-linked single-strand conformation polymorphism (F-PCR-SSCP) assay for the identification of Fasciola spp.

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Abstract

The present study aimed to establish a fluorescence-based polymerase chain reaction-linked single-strand conformation polymorphism (F-PCR-SSCP) assay for the identification of Fasciola spp. Based on the sequences of the second internal transcribed spacer (ITS-2) of the nuclear ribosomal DNA, we designed a set of genus-specific primers for the amplification of Fasciola ITS-2, with an estimated size of 140 bp. These primers were labelled by fluorescence dyes, and the PCR products were analyzed by capillary electrophoresis under non-denaturing conditions (F-PCR-SSCP). Capillary electrophoresis analysis of the fluorescence-labelled DNA fragments displayed three different peak profiles that allowed the accurate identification of Fasciola species: one single peak specific for either Fasciola hepatica or Fasciola gigantica and a doublet peak corresponding to the “intermediate” Fasciola. Validation of our novel method was performed using Fasciola specimens from different host animals from China, Spain, Nigeria, and Egypt. This F-PCR-SSCP assay provides a rapid, simple, and robust tool for the identification and differentiation between Fasciola spp.

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Acknowledgements

The authors are indebted to Ana L. García-Pérez and Ramón A. Juste (Sanidad Animal, Instituto Vasco de Investigación y Desarrollo Agrario, NEIKER), and Margarita Buades (Head of the Sección de Mataderos, Islas Baleares) for providing Fasciola samples and for their helpful comments. This project was funded by the programme “Ayudas a grupos de investigación” to RNM118 investigation group (Spain). XQZ is supported by the State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences. The experiments comply with the current laws of the countries in which the experiments were performed.

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Authors and Affiliations

  1. Estación Biológica de Doñana, Consejo Superior de Investigaciones Científicas (CSIC), Avda. Américo Vespucio s/n, 41092, Seville, Spain

    Samer Alasaad, Ramón C. Soriguer & Ana Píriz

  2. Department of Health Sciences, College of Arts and Sciences, Qatar University, Doha, Qatar

    Marawan Abu-Madi

  3. Parasitology Department, Faculty of Veterinary Medicine, Cairo University, Cairo, Arab Republic of Egypt

    Ahmed El Behairy

  4. Departamento de Patología Animal, Parasitología y Enfermedades Parasitarias, Facultad de Veterinaria, Universidad de Santiago de Compostela, 27071, Lugo, Spain

    Pablo Díez Baños

  5. Research Group Evolutionary Genetics, Leibniz-Institute for Zoo and Wildlife Research, Alfred-Kowalke-Str. 17, 10315, Berlin, Germany

    Joerns Fickel

  6. State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, CAAS, Lanzhou, Gansu Province, 730046, Peoples Republic of China

    Xing-Quan Zhu

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  1. Samer Alasaad
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  2. Ramón C. Soriguer
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  3. Marawan Abu-Madi
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  4. Ahmed El Behairy
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  5. Pablo Díez Baños
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  6. Ana Píriz
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  7. Joerns Fickel
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  8. Xing-Quan Zhu
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Correspondence to Samer Alasaad or Xing-Quan Zhu.

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Alasaad, S., Soriguer, R.C., Abu-Madi, M. et al. A fluorescence-based polymerase chain reaction-linked single-strand conformation polymorphism (F-PCR-SSCP) assay for the identification of Fasciola spp.. Parasitol Res 108, 1513–1517 (2011). https://doi.org/10.1007/s00436-010-2209-z

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