High-throughput capillary electrophoresis for the identification and differentiation of seven species of Eimeria from chickens.
A capillary electrophoretic approach has been evaluated for the identification of seven currently recognised species of Eimeria infecting chickens. The second internal transcribed spacer of ribosomal DNA is PCR-amplified from any of the seven species using a single set of oligonucleotide primers (one of which is fluorescently labelled). The amplicons are heat-denatured and subjected to capillary electrophoresis in a MegaBACE 1000 (Amersham). The chromatograms captured are stored electronically and then analysed using MegaBACE Fragment Profiler software. Using control DNA samples representing monospecific lines of Eimeria, specific peaks in the chromatograms were defined for the unequivocal identification of each of the seven species and their differentiation. Electrophoretic reading and analysis are carried out automatically, thus making it a time- and cost-effective method. This procedure should find applicability as a tool for the quality control of Eimeria vaccines, the monitoring of coccidiosis outbreaks and the high-throughput analysis of oocyst samples for epidemiological surveys.
Gasser
Robin B
RB
Department of Veterinary Science, The University of Melbourne, Werribee, Victoria, Australia. robinbg@unimelb.edu.au
Skinner
Rob
R
Fadavi
Reza
R
Richards
Grant
G
Morris
Genevieve
G
D000818:Animals; D001315:Australia; D002645:Chickens / Q000469:parasitology*; D003048:Coccidiosis / Q000469:parasitology / Q000662:veterinary*; D004275:DNA, Ribosomal / Q000032:analysis; D021903:DNA, Ribosomal Spacer / Q000032:analysis; D004539:Eimeria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D019075:Electrophoresis, Capillary / Q000379:methods*; D011201:Poultry Diseases / Q000469:parasitology*
D004275:DNA, Ribosomal; D021903:DNA, Ribosomal Spacer