title: Comparative evaluation of the nested ITS PCR against the 18S PCR-RFLP in a survey of bovine trypanosomiasis in Kwale County, Kenya.
abstract: We compared the nested internal transcribed spacer (ITS) PCR and the 18S PCR-RFLP (restriction-fragment length polymorphism) pan-trypanosome assays in a cross-sectional survey of bovine trypanosomiasis in 358 cattle in Kwale County, Kenya. The prevalence of trypanosomiasis as determined by the nested ITS PCR was 19.6% (70/358) and by 18S PCR-RFLP was 16.8% (60/358). Of the pathogenic trypanosomes detected, the prevalence of Trypanosoma congolense and Trypanosoma vivax was greater than that of Trypanosoma simiae The nested ITS PCR detected 83 parasite events, whereas the 18S PCR-RFLP detected 64; however, overall frequencies of infections and the parasite events detected did not differ between the assays (χ(2) = 0.8, df = 1, p > 0.05 and χ(2) = 2.5, df = 1, p > 0.05, respectively). The kappa statistic (0.8) showed good agreement between the tests. The nested ITS PCR and the 18S PCR-RFLP had comparable sensitivity, although the nested ITS PCR was better at detecting mixed infections (χ(2) = 5.4, df = 1, p < 0.05).
authors.affiliation: Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, Belgium (Odongo, Bekkele, Magez)Structural Biology Research Center (SBRC), Vlaams Instituut voor Biotechnologie (VIB), Brussels, Belgium (Odongo, Bekkele, Magez)Department of Biotechnical and Diagnostic Sciences, College of Veterinary Medicine, Animal Resources and Bio-security, Makerere University, Kampala, Uganda (Odongo)Interuniversity Programme Molecular Biology, Faculty of Science and Bio-engineering Sciences, Vrije Universiteit Brussel, Brussels, Belgium (Delespaux)Department of Animal Health and Production, Mount Kenya University, Thika, Kenya (Ngotho) sodongo@vub.ac.be.
authors.forename: Steven
authors.initials: S
authors.lastname: Odongo
authors.affiliation: Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, Belgium (Odongo, Bekkele, Magez)Structural Biology Research Center (SBRC), Vlaams Instituut voor Biotechnologie (VIB), Brussels, Belgium (Odongo, Bekkele, Magez)Department of Biotechnical and Diagnostic Sciences, College of Veterinary Medicine, Animal Resources and Bio-security, Makerere University, Kampala, Uganda (Odongo)Interuniversity Programme Molecular Biology, Faculty of Science and Bio-engineering Sciences, Vrije Universiteit Brussel, Brussels, Belgium (Delespaux)Department of Animal Health and Production, Mount Kenya University, Thika, Kenya (Ngotho).
authors.forename: Vincent
authors.initials: V
authors.lastname: Delespaux
authors.affiliation: Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, Belgium (Odongo, Bekkele, Magez)Structural Biology Research Center (SBRC), Vlaams Instituut voor Biotechnologie (VIB), Brussels, Belgium (Odongo, Bekkele, Magez)Department of Biotechnical and Diagnostic Sciences, College of Veterinary Medicine, Animal Resources and Bio-security, Makerere University, Kampala, Uganda (Odongo)Interuniversity Programme Molecular Biology, Faculty of Science and Bio-engineering Sciences, Vrije Universiteit Brussel, Brussels, Belgium (Delespaux)Department of Animal Health and Production, Mount Kenya University, Thika, Kenya (Ngotho).
authors.forename: Maina
authors.initials: M
authors.lastname: Ngotho
authors.affiliation: Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, Belgium (Odongo, Bekkele, Magez)Structural Biology Research Center (SBRC), Vlaams Instituut voor Biotechnologie (VIB), Brussels, Belgium (Odongo, Bekkele, Magez)Department of Biotechnical and Diagnostic Sciences, College of Veterinary Medicine, Animal Resources and Bio-security, Makerere University, Kampala, Uganda (Odongo)Interuniversity Programme Molecular Biology, Faculty of Science and Bio-engineering Sciences, Vrije Universiteit Brussel, Brussels, Belgium (Delespaux)Department of Animal Health and Production, Mount Kenya University, Thika, Kenya (Ngotho).
authors.forename: Serkalem Mindaye
authors.initials: SM
authors.lastname: Bekkele
authors.affiliation: Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Brussels, Belgium (Odongo, Bekkele, Magez)Structural Biology Research Center (SBRC), Vlaams Instituut voor Biotechnologie (VIB), Brussels, Belgium (Odongo, Bekkele, Magez)Department of Biotechnical and Diagnostic Sciences, College of Veterinary Medicine, Animal Resources and Bio-security, Makerere University, Kampala, Uganda (Odongo)Interuniversity Programme Molecular Biology, Faculty of Science and Bio-engineering Sciences, Vrije Universiteit Brussel, Brussels, Belgium (Delespaux)Department of Animal Health and Production, Mount Kenya University, Thika, Kenya (Ngotho).
authors.forename: Stefan
authors.initials: S
authors.lastname: Magez
mesh_terms: D000818:Animals; D002417:Cattle; D003430:Cross-Sectional Studies; D005260:Female; D007630:Kenya / Q000453:epidemiology; D008297:Male; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012150:Polymorphism, Restriction Fragment Length; D011237:Predictive Value of Tests; D015995:Prevalence; D014345:Trypanosoma / Q000235:genetics / Q000302:isolation & purification*; D014348:Trypanosoma congolense / Q000235:genetics / Q000302:isolation & purification; D016689:Trypanosoma vivax / Q000235:genetics / Q000302:isolation & purification; D014354:Trypanosomiasis, Bovine / Q000453:epidemiology* / Q000469:parasitology
keywords: 18S PCR-RFLP; Bovine trypanosomiasis; Kenya; Kwale County; nested ITS PCR