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Development of TaqMan-MGB based real-time PCR assay for detection of Babesia caballi
Pages: 54-58
Year: Issue:  1
Journal: Chinese Journal of Preventive Veterinary Medicine

Keyword:  Babesia caballireal-time PCR;
Abstract: To establish the rapid and sensitive assay for Babesia caballi detection, a TaqMan-MGB based real-time PCR was developed with a pair of primers and MGB probe designed according to the conserved sequence of the Bc-48 gene of B.caballi. Under the optimized reaction conditions, the established standard carve was linearity (R2=0.9996). The assay was specific to detect B.caballi, but had no amplification from the DNA of B.bigemina, Theileria. equi, T.annulata. and T.sergenti. The sensitivity of the method was 51.7 copies/μL, which was 100 times sensitive than that of conventional PCR. The reproducibility tests in inter-assay and intra-assay indicated that the coefficients of variation were both less than 1%. Moreover, a total of 90 clinical samples were tested by the assay and the conventional PCR, the positive rates was 53% and 30%, respectively. The assay established in this study was sensitive, specific and stable, which could be applied for B.caballi diagnosis and epidemiological investigation.
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