Abstract：
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 Theileria orientalis and Theileria annulata simultaneously by targeting 18 s rRna and MPSP (surface merozoite protein) genes, respectively. Fifty-five EDTA blood samples from clinically Theileria-suspected cows of three Theileria-endemic districts of Odisha were processed using acridine dye based fluorescent microscopy, Giemsa staining, and PCR. PCR revealed T. annulata and T. orientalis in 11/42 (26.11%) and 24/42 (57.14%) cases, respectively. Mixed infection due to both the Theileria 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 T. annulata and T. orientalis simultaneously. Acridine dye based fluorescent microscopy is comparatively easy and rapid method in detection of Thelieria spp.
摘要：
Theileriosis是热带地区牛的原生动物疾病，以生产损失和死亡的形式对奶农造成严重的经济损失。我们通过使用Taq-Man®探针设计并优化了多重实时PCR，通过靶向18srRNA和MPSP（表面裂殖子蛋白）基因同时检测和定量东方Theileria和环形Theileria，分别。使用基于a啶染料的荧光显微镜处理了来自奥里萨邦三个Theileria流行区的临床上怀疑Theileria的母牛的55份EDTA血液样本，Giemsa染色,和PCR。PCR在11/42例（26.11％）和24/42例（57.14％）中显示出环T.分别。由于Theileriaspp引起的混合感染。记录在7/42（16.66%）。与黄金标准测试（PCR）的比较，准确性,灵敏度,Giemsa染色的特异性分别为92.72、95.12和85.71％，吖啶橙染料的特异性分别为96.36、97.56和92.85％。使用Taq-Man探针的多重实时PCR同时检测了两种T.nulata和T.orientalis。基于acridine染料的荧光显微镜是一种相对简单，快速的检测Thelieriaspp的方法。
