Development and evaluation of a real-time polymerase chain reaction test for the detection of Theileria parva infections in Cape buffalo (Syncerus caffer) and cattle
Department of Veterinary Tropical Diseases, Faculty of Veterinary Science, University of Pretoria, Private Bag X04, Onderstepoort, 0110, South Africa; Department of Animal Health, Institute of Tropical Medicine, 155 Nationalestraat, Antwerp, B-2000, Belgium; Agricultural Research Council-Onderstepoort Veterinary Institute, Private Bag X5, Onderstepoort, 0110, South Africa; Department of Statistics, School of Mathematical Science, University of Pretoria, Pretoria, 0002, South Africa
Corridor disease, caused by the tick-borne protozoan parasite Theileria parva, is a controlled disease in South Africa. The Cape buffalo is the reservoir host and uninfected buffalo have become sought-after by the game industry in South Africa, particularly for introduction into Corridor disease-free areas. A real-time polymerase chain reaction (PCR) test for detection of T. parva DNA in buffalo and cattle was developed to improve the sensitivity and specificity of the official diagnostic test package in South Africa. Oligonucleotide primers and hybridization probes were designed based on the 18S ribosomal RNA (rRNA) gene. Amplification of control DNA using Theileria genus-specific primers resulted in detection of T. taurotragi and T. annulata, in addition to T. parva. A T. parva-specific forward primer was designed which eliminated amplification of all other Theileria species, except for Theileria sp. (buffalo); however only the T. parva product was detected by the T. parva-specific hybridization probe set. The real-time PCR assay requires less time to perform, is more sensitive than the other molecular assays previously used in T. parva diagnostics and can reliably detect the parasite in carrier animals with a piroplasm parasitaemia as low as 8.79 × 10 -4%. © 2008 Elsevier B.V. All rights reserved.
animal experiment; animal parasitosis; article; buffalo; cattle; controlled study; diagnostic test; gene amplification; intermethod comparison; methodology; nonhuman; nucleotide sequence; real time polymerase chain reaction; RNA gene; sensitivity and specificity; Theileria; Theileria annulata; Theileria parva; theileria taurotragi; Animals; Buffaloes; Cattle; Disease Reservoirs; DNA, Protozoan; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Sensitivity and Specificity; Theileria parva; Theileriasis; Animalia; Bos; Ixodida; Protozoa; Syncerus caffer; Syncerus caffer caffer; Theileria; Theileria annulata; Theileria parva; Theileria sp.; Theileria taurotragi