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A specific primer pair for the diagnosis and identification of Acanthamoeba astronyxis by random amplified polymorphic DNA-polymerase chain reaction
Authors:Ortega-Rivas A  Lorenzo-Morales J  Martínez E  Villa M  Clavel A  Valladares B  del Castillo A
Institution:Faculty of Farmacy, Department of Parasitology, University of La Laguna. Avda. Francisco Sánchez s/n, La Laguna CP 38271, Canary Islands, Spain. aortega@ull.es
Abstract:Random amplified polymorphic DNA (RAPD) is a useful tool for species identification. The obtained band patterns can be used for specific primer pair design that is useful for species identification. In this study, a distinctive 485-bp band in Acanthamoeba astronyxis band patterns was found, using the OPC20 primer (ACTTCGCCAC). The band specificity was confirmed by hybridization, using it as a probe, against all OPC20 amplifications from different Acanthamoeba species. Once the fragment was sequenced, we used it to design a specific primer pair that was useful for the identification of different isolates as A. astronyxis species.
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