To detect the Babesia bovis, a real-time PCR method was established with a pair of primers and a TaqMan probe designed according to the conserved sequence of 18S rRNA genes in GenBank. The results showed that: the limit detection of real-time PCR was about 1.31×101 copies/μL for the target gene which was 1,000 times higher sensitivity than conventional PCR, and no cross-reaction with other piroplasmasida in cattle. The reproducibility tests in inter-assay and intra-assay indicated that the coefficients of variation were less than 3%. A total of 23 clinical samples were tested by the real-time PCR comparing with conventional PCR, and positive rates were 52.17% (12/23) and 30.43% (7/23), respectively. The establishment of the real-time PCR method for detection of bovine Babesia was sensitive, specific and rapid detection technology, which is provided a assay for the tick-borne bovine piroplasmosis detection.%为建立牛巴贝斯虫(B.bovis)的TaqMan实时荧光PCR检测方法,本研究根据GenBank中B.bovis的18S rRNA基因保守序列,设计引物和TaqMan探针,通过优化反应体系,建立检测B.bovis的实时荧光PCR方法.试验结果表明:实时荧光PCR对靶基因的最低检测值为1.31×101 copies/μL,比常规PCR的敏感性高1 000倍；而且与牛的其他血液原虫无交叉反应；组内及组间重复性试验的变异系数均小于3％,具有良好的重复性；在23份被检样品中,实时荧光PCR和常规PCR的检出率分别为52.17％和30.43％.该检测方法的建立为B.bovis的检测提供了一种快速、敏感、特异的技术手段.
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