期刊
VETERINARY PARASITOLOGY
卷 162, 期 3-4, 页码 200-206出版社
ELSEVIER
DOI: 10.1016/j.vetpar.2009.03.040
关键词
Babesia; Theileria; Quantitative PCR; Molecular diagnostic; TaqMan probes; FRET probes
资金
- EC [INCO-MEDLABAB 003691]
This paper reports two new quantitative PCR (qPCR) assays, developed in an attempt to improve the detection of bovine piroplasmids. The first of these techniques is a duplex TaqMan assay for the simultaneous diagnosis of Babesia bovis and B. bigemina. This technique is ideal for use in South America where bovids harbour no theilerids. The second technique, which is suitable for the diagnosis of both babesiosis and theileriosis worldwide, involves fluorescence resonance energy transfer (FRET) probes. In FRET assays, Babesia bovis, B. divergens, Babesia sp. (B. major or B. bigemina), Theileria annae and Theileria sp. were all identifiable based on the melting temperatures of their amplified fragments. Both techniques provided linear calibration curves over the 0.1 fg/mu l to 0.01 ng/mu l DNA range. The assays showed good sensitivity and specificity. To assess their performance, both procedures were compared in two separate studies: the first was intended to monitor the experimental infection of calves with B. bovis and the second was a survey where 200 bovid/equine DNA samples from different countries were screened for piroplasmids. Comparative studies showed that duplex TaqMan qPCR was more sensitive than FRET qPCR in the detection of babesids. (C) 2009 Elsevier B.V. All rights reserved.
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