Establishment and application of TaqMan real-time PCR method for detection of Theileria annulata resistant to buparvaquone

作者：Shuxiao Su, Shuaiyang Zhao, Junlong Liu, Chuhan Zhang, Haohan Zhu, Guiquan Guan, Hong Yin, Jianxun Luo · 发表于：Veterinary Parasitology · 年份：2024 · DOI：10.1016/j.vetpar.2024.110183 · 被引用次数：5 · 研究领域：Vector-borne infectious diseases、Insect and Pesticide Research、Toxin Mechanisms and Immunotoxins

Tropical theileriosis is a tick-borne disease that caused by Theileria annulata, and leads to substantial economic impact in endemic area. Distinguishes to other piroplasms, Theileria is the only eukaryotic parasite could transform mammalian leukocytes. At present, buparvaquone is the most effective drug used for treatment of Theileria infection. However, frequently reported of failure treatment with buparvaquone for some T. annulata isolates. Mutation of TaPIN1 was reported to be the direct reason for failure of buparvaquone treatment. Through in vitro culture, a T. annulata isolate with a TaPIN1 mutation that is similar to the reported strain was recently identified in China. In order to understand the distribution of Theileria with mutation of TaPIN1 in China, here we developed a TaqMan probe-based real-time PCR technology to detect the mutated TaPIN1 gene. The specificity, sensitivity and reproducibility of the established TaqMan Real-time PCR method were evaluated, and field cattle blood samples collected from Xinjiang Uyghur Autonomous Region were used to test its application. Among 1683 samples, 335 samples were confirmed positive for T. annulata by traditional PCR method and 34 samples were positive for buparvaquone-resistant. The TaPIN1 gene of those 34 samples was sequenced and analyzed with the published gene sequences from NCBI database. The results showed that the sequence obtained from the present study has good consistency with those published sequences. In con...