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Title

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

AuthorsAlasaad, Samer ; Soriguer, Ramón C. ; Abu-Madi, Marawan; El Behairy, Ahmed; Díez Baños, Pablo; Píriz, Ana ; Fickel, Joerns; Sánchez, Antonio
Issue DateJun-2011
PublisherSpringer
CitationParasitol Res (2011) 108:1513–1517
AbstractThe 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 electropho- resis 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.
Publisher version (URL)http://dx.doi.org/10.1007/s00436-010-2209-z
URIhttp://hdl.handle.net/10261/49508
DOI10.1007/s00436-010-2209-z
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