Authors: David O Odongo Jack D Sunter Henry K Kiara Robert A Skilton Richard P Bishop
Publish Date: 2009/11/10
Volume: 106, Issue: 2, Pages: 357-
Abstract
Theileria parva causes East Coast fever an economically important disease of cattle in subSaharan Africa We describe a nested polymerase chain reaction nPCR assay for the detection of T parva DNA in cattle blood spotted onto filter paper using primers derived from the T parvaspecific 104kDa antigen p104 gene The sensitivity of this assay was compared to a previously described p104based PCR and also the reverse line blot RLB technique using serial dilutions of blood from a calf with known T parva piroplasm parasitaemia The relative sensitivities of the three assays were 04 14 and 4 parasites/µl corresponding to blood parasitaemias of 92 × 10−6 28 × 10−5 and 83 × 10−5 respectively The three assays were applied to samples from two calves infected with the T parva Muguga stock Parasite DNA was consistently detectable by the two p104 PCR assays until 48 and 82 days postinfection respectively and thereafter sporadically RLB detected parasite DNA in the two infected calves until days 43 and 45 Field samples from 151 Kenyan cattle exhibited 377 positivity for T parva by regular p104 PCR and 423 positivity using p104 nPCR Among 169 cattle blood samples from Southern Sudan 36 were positive for T parva using nPCR The nPCR assay represents a highly sensitive tool for detection and monitoring of asymptomatic carrier state infections of T parva in the blood of cattleThis study was partially supported by a financial grant from the International Fund on Agricultural Development IFAD We thank Thomas Njoroge for his skilled technical assistance and to Chris Oura for help in the initial setting up of the RLB assay This is ILRI publication no 200907
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