CHARACTERIZATION OF SPANISH TRICHINELLA ISOLATES BY RANDOM AMPLIFIED POLYMORPHIC DNA (RAPD)

The random amplified polymorphic DNA (RAPD) assay was used to find mol ecular markers able to distinguish Trichinella spiralis from T. britov i, the two recognized Spanish Trichinella species. Fourteen Spanish Tr ichinella isolates, as well as reference Trichinella isolates represen ting the five species T. spiralis (T1), T. nativa (T2), T. britovi (T3 ), T. pseudospiralis (T4) and T. nelsoni (T7) and the three other taxa Trichinella T5, Trichinella T6 and Trichinella T8 of the genus, were characterized by RAPD using both purified and crude DNAs from infectiv e muscle larvae (ML) and seven arbitrary primers. Three primers yielde d diagnostic RAPD markers for the Spanish T. spiralis and T. britovi i solates as well as for the Trichinella reference isolates analysed, an d in the case of crude DNAs the results were obtained in few hours. In addition, the species-specificity of the diagnostic RAPD markers from Spanish Trichinella isolates was studied by cross-hybridization assay s. These assays confirmed that the selected diagnostic DNA fragments w ere not species-specific, but showed potential differences in the copy number among the examined Trichinella genetic clusters.