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    Comparison of the performance of three PCR assays for the detection and differentiation of Theileria orientalis genotypes

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    17
    Author
    Perera, PK; Gasser, RB; Pulford, DJ; Stevenson, MA; Firestone, SM; McFadden, AMJ; Jabbar, A
    Date
    2015-03-31
    Source Title
    Parasites and Vectors
    Publisher
    BIOMED CENTRAL LTD
    University of Melbourne Author/s
    Firestone, Simon; Gasser, Robin; Jabbar, Abdul; Stevenson, Mark; Perera, Piyumali
    Affiliation
    Veterinary Biosciences
    Melbourne School of Population and Global Health
    Metadata
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    Document Type
    Journal Article
    Citations
    Perera, P. K., Gasser, R. B., Pulford, D. J., Stevenson, M. A., Firestone, S. M., McFadden, A. M. J. & Jabbar, A. (2015). Comparison of the performance of three PCR assays for the detection and differentiation of Theileria orientalis genotypes. PARASITES & VECTORS, 8 (1), https://doi.org/10.1186/s13071-015-0812-7.
    Access Status
    Open Access
    URI
    http://hdl.handle.net/11343/260183
    DOI
    10.1186/s13071-015-0812-7
    Abstract
    BACKGROUND: Oriental theileriosis is a tick-borne disease of bovines caused by the members of the Theileria orientalis complex. Recently, we developed a multiplexed tandem (MT) PCR to detect, differentiate and quantitate four genotypes (i.e., buffeli, chitose, ikeda and type 5) of T. orientalis. In this study, we used MT PCR to assess the prevalence and infection intensity of four T. orientalis genotypes in selected cattle herds that experienced oriental theileriosis outbreaks in New Zealand, and compared the sensitivities and specificities of MT PCR, PCR-high resolution melting (PCR-HRM) and a TaqMan qPCR. METHODS: MT PCR, PCR-HRM analysis for T. orientalis and a TaqMan qPCR assay for ikeda genotype were employed to test 154 and 88 cattle blood samples from North (where oriental theileriosis outbreaks had occurred; designated as Group 1) and South (where no outbreaks had been reported; Group 2) Islands of New Zealand, respectively. Quantitative data from MT PCR assay were analyzed using generalized linear model and paired-sample t-test. The diagnostic specificity and sensitivity of the assays were estimated using a Bayesian latent class modeling approach. RESULTS: In Group 1, 99.4% (153/154) of cattle were test-positive for T. orientalis in both the MT PCR and PCR-HRM assays. The apparent prevalences of genotype ikeda in Group 1 were 87.6% (134/153) and 87.7% (135/154) using the MT PCR and Ikeda TaqMan qPCR assays, respectively. Using the MT PCR test, all four genotypes of T. orientalis were detected. The infection intensity estimated for genotype ikeda was significantly higher (P = 0.009) in severely anaemic cattle than in those without anaemia, and this intensity was significantly higher than that of buffeli (P < 0.001) in the former cattle. Bayesian latent class analysis showed that the diagnostic sensitivities (97.1-98.9%) and specificities (96.5-98.9%) of the three PCR assays were very comparable. CONCLUSION: The present findings show the advantages of using the MT PCR assay as a useful tool for in-depth epidemiological and transmission studies of T. orientalis worldwide.

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