Evaluation of a real-time PCR test for the detection and discrimination of Theileria species in the African buffalo (Syncerus caffer)

dc.contributor.authorChaisi, Mamohale E.
dc.contributor.authorJanssens, Michiel E.
dc.contributor.authorVermeiren, Lieve
dc.contributor.authorOosthuizen, Marinda C.
dc.contributor.authorCollins, Nicola E.
dc.contributor.authorGeysen, Dirk
dc.date.accessioned2013-11-12T09:09:09Z
dc.date.available2013-11-12T09:09:09Z
dc.date.issued2013-10-17
dc.description.abstractA quantitative real-time PCR (qPCR) assay based on the cox III gene was evaluated for the simultaneous detection and discrimination of Theileria species in buffalo and cattle blood samples from South Africa and Mozambique using melting curve analysis. The results obtained were compared to those of the reverse line blot (RLB) hybridization assay for the simultaneous detection and differentiation of Theileria spp. in mixed infections, and to the 18S rRNA qPCR assay results for the specific detection of Theileria parva. Theileria parva, Theileria sp. (buffalo), Theileria taurotragi, Theileria buffeli and Theileria mutans were detected by the cox III assay. Theileria velifera was not detected from any of the samples analysed. Seventeen percent of the samples had non-species specific melting peaks and 4.5% of the samples were negative or below the detection limit of the assay. The cox III assay identified more T. parva and Theileria sp. (buffalo) positive samples than the RLB assay, and also detected more T. parva infections than the 18S assay. However, only a small number of samples were positive for the benign Theileria spp. To our knowledge T. taurotragi has never been identified from the African buffalo, its identification in some samples by the qPCR assay was unexpected. Because of these discrepancies in the results, cox III qPCR products were cloned and sequenced. Sequence analysis indicated extensive inter- and intra-species variations in the probe target regions of the cox III gene sequences of the benign Theileria spp. and therefore explains their low detection. The cox III assay is specific for the detection of T. parva infections in cattle and buffalo. Sequence data generated from this study can be used for the development of a more inclusive assay for detection and differentiation of all variants of the mildly pathogenic and benign Theileria spp. of buffalo and cattle.en
dc.description.librarianam2013en
dc.description.librarianab2013
dc.description.sponsorshipThis work was part of a PhD project that was funded by the South African National Research Foundation (NRF ICD2006072000009) and UP Research Development Programme.en
dc.description.urihttp://www.plosone.orgen
dc.identifier.citationChaisi ME, Janssens ME, Vermeiren L, Oosthuizen MC, Collins NE, et al. (2013) Evaluation of a Real-Time PCR Test for the Detection and Discrimination of Theileria Species in the African Buffalo (Syncerus caffer). PLoS ONE 8(10): e75827. DOI: 10.1371/journal.pone.0075827en
dc.identifier.issn1932-6203
dc.identifier.other10.1371/journal.pone.0075827
dc.identifier.other7004592997
dc.identifier.other7103250386
dc.identifier.otherN-8706-2014
dc.identifier.otherO-6342-2014
dc.identifier.urihttp://hdl.handle.net/2263/32374
dc.language.isoenen
dc.publisherPublic Library of Scienceen
dc.relation.requiresAdobe Acrobat Readeren
dc.rights© 2013 Chaisi et al. This is an open-access article distributed under the terms of the Creative Commons Attribution Licenseen
dc.subjectQuantitative real-time polymerase chain reaction (qRT-PCR)en
dc.subjectAfrican buffalo (Syncerus caffer)en
dc.subject.lcshTheileriaen
dc.titleEvaluation of a real-time PCR test for the detection and discrimination of Theileria species in the African buffalo (Syncerus caffer)en
dc.typeArticleen

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