Development and application of Real-time PCR assay for detection of Theilera annulata infection in cattle in Xinjiang Uygur Autonomous Region, China
- Author:
Guo, Q.Y., Cao, W.L., Wang, B.J., Chahan
- Publication Type:Journal Article
- From:Tropical Biomedicine
2016;33(1):27-34
- CountryMalaysia
- Language:English
-
Abstract:
Theileria annulata (T. annulata), the causative agent of tropical theileriosis, is a
protozoan parasite that also causes lymphoproliferative diseases in cattle. Development of
reliable and fast methods are necessary in the epidemiological investigation of T. annulata in
ticks and animals. Real-time PCR possesses merits of rapidity, accuracy, reliability, automation
and ease of standardization, which is widely used for the detection of blood borne parasites.
In this study, species-specific primers and TaqMan probe were designed on the basis of the
18s rRNA gene sequence of T. annulata, and the real-time PCR assay was developed by
optimizing the reaction parameter. The performance of real-time PCR was assessed by testing
47 blood samples from cattle and comparing with the results from conventional PCR. The
results show that this real-time PCR assay could specifically detect 10 copies DNA of T.
annulata, which is 10-fold sensitivity more than conventional PCR. No cross-reactions were
observed with Babesia bovis, Babesia bigemina, Trypanosoma evansi and Theileria equi. Of
the 47 field samples collected from Xinjiang Uygur Autonomous Region, China, 36.17% were
detected by real-time PCR, and 25.53% were found positive for T. annulata infection by
conventional PCR. These results indicated that the real-time PCR assay is a useful approach
for detecting T. annulata infections and has potential as an alternative tool for ecological
and epidemiological surveillance of ovine theileriosis.
- Full text:P020160419511970275282.pdf
