A fluorescence-based polymerase chain reaction-linked single-strand conformation polymorphism (F-PCR-SSCP) assay for the identification of Fasciola spp
Author | Alasaad, Samer |
Author | Soriguer, Ramón C |
Author | Abu-Madi, Marawan |
Author | El Behairy, Ahmed |
Author | Baños, Pablo Díez |
Author | Píriz, Ana |
Author | Fickel, Joerns |
Author | Zhu, Xing-Quan |
Available date | 2017-12-07T09:46:56Z |
Publication Date | 2011-06-01 |
Publication Name | Parasitology Research |
Identifier | http://dx.doi.org/10.1007/s00436-010-2209-z |
Citation | Alasaad, S., Soriguer, R.C., Abu-Madi, M. et al.(2011), Parasitol Res, 108(6), pp. 1513–1517. |
ISSN | 0932-0113 |
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. |
Sponsor | Funded by the programme “Ayudas a grupos de investigación” to RNM118 investigation group (Spain). |
Language | en |
Publisher | Springer Verlag |
Subject | F-PCR-SSCP Fasciola spp |
Type | Article |
Pagination | 1513–1517 |
Issue Number | 6 |
Volume Number | 108 |
ESSN | 1432-1955 |
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