Comparative evaluation of diagnostic methods for detection of Theileria spp. in cows.

Theileriosis is a hemoprotozoan illness of cattle in tropical regions that poses a severe economic loss to dairy farmers in the form of production loss and mortality. We designed and optimized a multiplex real-time PCR by using Taq-Man probe for detection and quantification of and simultaneously by targeting 18 s rRna and MPSP (surface merozoite protein) genes, respectively. Fifty-five EDTA blood samples from clinically -suspected cows of three -endemic districts of Odisha were processed using acridine dye based fluorescent microscopy, Giemsa staining, and PCR. PCR revealed and in 11/42 (26.11%) and 24/42 (57.14%) cases, respectively. Mixed infection due to both the spp. was recorded in 7/42 (16.66%). On comparison with gold standard test (PCR), the accuracy, sensitivity, and specificity were 92.72, 95.12, and 85.71% for Giemsa staining and 96.36, 97.56, and 92.85% for acridine orange dye. Multiplex real time PCR using Taq-Man probe detected two species of and simultaneously. Acridine dye based fluorescent microscopy is comparatively easy and rapid method in detection of spp.