Detection of Theileria ovis in naturally infected sheep by nested PCR.
A nested polymerase chain reaction (PCR) for the detection of Theileria ovis in sheep using oligonucleotide primers designed from the small subunit ribosomal RNA (SSU rRNA) gene sequence of T. ovis from sheep in eastern Turkey is described. A 398-bp DNA fragment was specifically amplified from blood samples from sheep, naturally infected with T. ovis. No PCR products resulted from T. lestoquardi, T. annulata, T. parva, T. buffeli and Babesia spp. DNA using these specific primers. The sensitivity of the nested PCR for T. ovis, which was assessed showed that one infected cell in 10(7) sheep erythrocytes, equivalent to a blood parasitemia of 0.00001%, could be detected. This is more sensitive than examining 200 fields under light microscopy. In addition, of the 124 field samples obtained from sheep in eastern Turkey tested, 19.35% (24/124) were positive for the presence of Theileria spp. by microscopic examination compared to 54.03% (67/124) positive for T. ovis by nested PCR. The primer pairs described in this study will be useful for epidemiological studies on ovine theileriosis and for discrimination between T. lestoquardi and T. ovis infections in sheep.
Altay
Kursat
K
Department of Parasitology, Faculty of Veterinary Medicine, University of Firat, 23119 Elazig, Turkey.
Dumanli
Nazir
N
Holman
Patricia J
PJ
Aktas
Munir
M
D000818:Animals; D017931:DNA Primers; D016054:DNA, Protozoan / Q000032:analysis; D004275:DNA, Ribosomal / Q000032:analysis; D004912:Erythrocytes / Q000469:parasitology; D008853:Microscopy / Q000662:veterinary; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis* / Q000453:epidemiology; D016794:Theileria / Q000302:isolation & purification*; D013801:Theileriasis / Q000175:diagnosis* / Q000453:epidemiology; D014421:Turkey / Q000453:epidemiology
D017931:DNA Primers; D016054:DNA, Protozoan; D004275:DNA, Ribosomal