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A nested PCR assay exhibits enhanced sensitivity for detection of Theileria parva infections in bovine blood samples from carrier animals

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Abstract

Theileria parva causes East Coast fever, an economically important disease of cattle in sub-Saharan 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. parva-specific 104-kDa antigen (p104) gene. The sensitivity of this assay was compared to a previously described p104-based 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 0.4, 1.4 and 4 parasites/µl corresponding to blood parasitaemias of 9.2 × 10−6%, 2.8 × 10−5% and 8.3 × 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 post-infection, 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 37.7% positivity for T. parva by regular p104 PCR and 42.3% 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 cattle.

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Acknowledgements

This 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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Authors and Affiliations

  1. The International Livestock Research Institute, P.O. Box 30709, Nairobi, Kenya

    David O. Odongo, Henry K. Kiara & Richard P. Bishop

  2. School of Biological Sciences, University of Nairobi, Nairobi, Kenya

    David O. Odongo

  3. Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge, CB2 1GA, UK

    Jack D. Sunter

  4. Biosciences Eastern and Central Africa, P.O. Box 30709, Nairobi, Kenya

    Robert A. Skilton

Authors
  1. David O. Odongo
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  2. Jack D. Sunter
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  3. Henry K. Kiara
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  4. Robert A. Skilton
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  5. Richard P. Bishop
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Correspondence to David O. Odongo.

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Odongo, D.O., Sunter, J.D., Kiara, H.K. et al. A nested PCR assay exhibits enhanced sensitivity for detection of Theileria parva infections in bovine blood samples from carrier animals. Parasitol Res 106, 357–365 (2010). https://doi.org/10.1007/s00436-009-1670-z

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  • DOI: https://doi.org/10.1007/s00436-009-1670-z

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