Abstract
As an alternative to morphological speciation, DNA probes have been developed to identifyEimeria crandallis andE. ovinoidalis, the species associated with ovine coccidiosis. Coccidia-free lambs were infected with pure cultures ofE. crandallis andE. ovinoidalis. Oocysts separated from faeces were sporulated and treated to release sporozoites from which DNA was purified. Repetitive DNA sequences were identified by hybridisation and cloned. The specificity of selected clones was tested by hybridisation with dot blots of four other ovineEimeria species, a mixture of caprineEimeria species andE. acervulina of the chicken. The probes allowed the detection of DNA equivalent to that present in approximately 1,500 oocysts. To increase the sensitivity of the test, a polymerase chain reaction (PCR) approach was adopted, using oligonucleotide primers specific for the cloned repeats. By this method, less than 10 oocysts ofE. ovinoidalis could be reliably detected. In addition to the species-specific probes, other highly repetitive DNA sequences fromE. crandallis andE. ovinoidalis were cloned and sequenced; these repeats hybridised to all ovine, caprine and avianEimeria species tested and contained tandem CGA and putative telomeric repeats.
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Berriatua, E., Gibson, W.C. & Morgan, K.L. Development of DNA probes for the ovine Eimeria speciesE. crandallis andE. ovinoidalis . Parasitol Res 81, 222–229 (1995). https://doi.org/10.1007/BF00937113
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DOI: https://doi.org/10.1007/BF00937113