Use of mitochondrial RNA genes for the differentiation of fourTrichinellaspecies by multiplex PCR amplification

Abstract

AbstractUntil now, four species of theTrichinellagenus have been identified in Europe:Trichinella spiralis,T. nativa,T. britoviandT. pseudospiralis. The aim of this work was to establish a sound polymerase chain reaction (PCR)-based method to differentiate these four species using mitochondrial rDNA as a reliable genetic marker and to evaluate the sensitivity of this method. Full-length DNA sequences coding for the small and large mitochondrial rRNA (mt-rrnSand mt-rrnL) of the four species are described. A multiplex PCR was designed and successfully tested on 24 European isolates. As few as one larva, or 100 pg of genomic DNA was detected, providing equivalent sensitivity to previously described PCR methods. The PCR-based method of mitochondrial rDNA amplification was thereby established as a sensitive and reproductive diagnostic method for the four EuropeanTrichinellaspecies.