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 transcribed spacer (ITS-2) was amplified from genomic DNA by PCR, digested separately 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 approximately 70 to 620 bp on agarose gels, with differences in size and number of bands among species, but no apparent variation within a species. The PCR-RFLP analysis of ITS-2 amplicons on denaturing gels gave characteristic profiles for 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.
Woods
W G
WG
Department of Veterinary Science, The University of Melbourne, 250 Princes Highway, Werribee, Victoria, Australia.
Whithear
K G
KG
Richards
D G
DG
Anderson
G R
GR
Jorgensen
W K
WK
Gasser
R B
RB
D000818:Animals; D001315:Australia; D002645:Chickens / Q000469:parasitology*; D003048:Coccidiosis / Q000469:parasitology / Q000662:veterinary*; D017931:DNA Primers / Q000737:chemistry; D021903:DNA, Ribosomal Spacer / Q000737:chemistry*; D004277:DNA, Single-Stranded / Q000737:chemistry; D004539:Eimeria / Q000737:chemistry / Q000145:classification / Q000235:genetics*; D004587:Electrophoresis, Agar Gel / Q000662:veterinary; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D005243:Feces / Q000469:parasitology; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012150:Polymorphism, Restriction Fragment Length; D011201:Poultry Diseases / Q000469:parasitology*; D013047:Specific Pathogen-Free Organisms
D017931:DNA Primers; D021903:DNA, Ribosomal Spacer; D004277:DNA, Single-Stranded