Molecular identification of equine strongyles by a Reverse Line Blot hybridization assay
Small and large strongyles (Nematoda, Strongylidae) are horse parasites of primary importance worldwide. More that 50 species are recognized, which can be microscopically identified on the basis of tails and heads of the adult stages. However, their identification can be challenging and the differentiation of the immature stages is almost impossible. The characterization of the intergenic spacers (IGS) of the ribosomal DNA (rDNA) recently provided powerful genetic markers for the molecular identification of six small strongyles. In this work the IGS of eight species of small strongyles and of three species of large strongyles from different geographical areas were characterized. Species-specific probes designed in the strongyle consensus IGS regions were evaluated in a Reverse Line Blot (RLB) Hybridization for their ability to simultaneously identify the 11 species of strongyles. The probes hybridized specifically with the IGS of the strongyles included in the assay, whose conditions were validated by varying incubation time and temperatures, and probes concentration. The RLB herein presented allows the simultaneous molecular differentiation of 11 equine strongyles species irrespective of their life cycle stage, thus providing a powerful tool for faecal diagnosis of horse strongylosis and biological and epidemiological studies of this infection.
Full conference title:
158th Meeting of the Society for General Microbiology