Molecular diagnostic procedures using RT PCR to alleviate taxonomic impediments of parasite species segregation

Abstract

The taxonomic impediments in taxa differentiation have been overcome by the application of advanced techniques using real time PCR methods PCR product detection and identification of helminth species is performed with a variety of fluorescence chemistries The measurement of melting temperatures Tms of amplicons after completion of PCR facilitated precise diagnosis of species of particular helminth The technique is reliable as the Tms of the specific amplicons are sufficiently different and constant to illustrate differentiation The phylogeny of Contrcaecum sp from a marine fish Sillago sihama is established with report of Tms C using melting curve analysis real time PCR in the present investigation with simultaneous segregation of Anisakis sp as a separate taxon from the coexisting species Johnius dussumieri whose genotypes have been reported at the GenBank