Single-strand restriction fragment length polymorphism analysis of the second internal transcribed spacer (ribosomal DNA) for six species of Eimeria from chickens in Australia

Species of Eimeria from chickens from Australia were characterised using a polymerase chain reaction-linked restriction fragment length polymorphism ( PCR-RFLP) approach. The ribosomal DNA region spanning the second internal t ranscribed spacer (ITS-2) was amplified from genomic DNA by PCR, digested s eparately with three restriction endonucleases (CfoI, Sau3AI and TaqI) and the fragments separated by denaturing gel electrophoresis. The PCR products amplified from the six species varied from similar to 70 to 620 bp on agar ose gels, with differences in size and number of bands among species, but n o apparent variation within a species. The PCR-RFLP analysis of ITS-2 ampli cons on denaturing gels gave characteristic profiles fbr individual species (except for minor variation in profiles within some species). The results indicate that ITS-2 contains useful genetic markers for the identification of six Eimeria species occurring in Australia. (C) 2000 Australian Society for Parasitology Inc. Published by Elsevier Science Ltd. All rights reserve d.