IDENTIFICATION OF PARASITIC NEMATODES BY PCR-SSCP OF ITS-2 RDNA

Fourteen species of parasitic nematodes (order Strongylida) were chara cterized using a polymerase chain reaction-linked single strand confor mation polymorphism technique (PCR-SSCP). The rDNA region spanning the second internal transcribed spacer (ITS-2) was amplified from parasit e DNA by PCR. The PCR products were then denatured and subjected to el ectrophoresis on a nondenaturing gel matrix. PCR-SSCP of the single st randed ITS-2 molecules generated characteristic and reproducible patte rns for each species, and allowed the rapid delineation of all of the 14 species in one step. The method also allowed the display of variati on in patterns within some species between different geographical isol ates. These findings demonstrate the usefulness of PCR-SSCP of ITS-2 f or the rapid identification of nematode species and indicate its poten tial for resolving variation in the ITS-2 sequence within a species. ( C) 1997 Academic Press Limited.