Tian Zhancheng, Du Junzheng, Yang Jifei, Liu Aihong, Liu Xiaocui, Liu Guangyuan, Yin Hong
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, 730046, P. R. China.
Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, 225009, P. R. China.
Parasit Vectors. 2015 Sep 17;8:475. doi: 10.1186/s13071-015-1085-x.
Bovine babesiosis and theileriosis is an important hemoprotozoal disease in cattles and yaks in tropical and subtropical regions leading to significant economic losses. In the field, the risk of co-infection between the bovine Babesia and Theileria species is very high. Thus, it is necessary to develop a simple, accurate, rapid and cost-effective method for large-scale epidemic investigation, in particular for the detection of co-infection in field.
In this study, DNA sequences of a ribosomal protein S8 (RPS8) gene from eight species of cattle piroplasms in China were used to develop a species-specific PCR-RFLP diagnostic tool. The eight Theileria and Babesia species could be differentiated by digesting the RPS8 PCR product with Mbo I.
The sensitivity of the PCR assays was 0.1 pg DNA for Babesia species but 1 pg DNA for Theileria species. The clearly different size of the PCR-RFLP products allowed for a direct discrimination between eight bovine Theileria and Babesia species (T. annulata, T. sinensis, T. sergenti, B. ovata, B. bovis, B. bigemina, B. major and Babesia species Kashi isolate).
Our results indicated that the established method based on the RPS8 gene was a reliable molecular diagnostic tool for the simultaneous detection and identification of bovine Babesia and Theileria species in China, which could be applicable for the survey of parasite dynamics, epidemiological studies as well as prevention and control of the disease.
牛巴贝斯虫病和泰勒虫病是热带和亚热带地区牛和牦牛的一种重要血液原虫病，会导致重大经济损失。在野外，牛巴贝斯虫和泰勒虫物种之间的共同感染风险非常高。因此，有必要开发一种简单、准确、快速且经济高效的方法用于大规模疫情调查，特别是用于检测野外的共同感染。
在本研究中，利用中国八种牛梨形虫的核糖体蛋白S8（RPS8）基因的DNA序列开发了一种物种特异性PCR-RFLP诊断工具。通过用Mbo I消化RPS8 PCR产物，可以区分这八种泰勒虫和巴贝斯虫物种。
PCR检测对巴贝斯虫物种的灵敏度为0.1 pg DNA，而对泰勒虫物种为1 pg DNA。PCR-RFLP产物明显不同的大小使得能够直接区分八种牛泰勒虫和巴贝斯虫物种（环形泰勒虫、中华泰勒虫、瑟氏泰勒虫、卵形巴贝斯虫、牛巴贝斯虫、双芽巴贝斯虫、大巴贝斯虫和喀什分离的巴贝斯虫物种）。
我们的结果表明，基于RPS基因建立的方法是一种可靠的分子诊断工具，可用于同时检测和鉴定中国的牛巴贝斯虫和泰勒虫物种，适用于寄生虫动态监测、流行病学研究以及疾病的预防和控制。
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