State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping, Yanchangbu, Lanzhou, Gansu Province 730046, People's Republic of China.
Vet Parasitol. 2013 Oct 18;197(1-2):354-9. doi: 10.1016/j.vetpar.2013.04.033. Epub 2013 Apr 26.
Ovine babesiosis and theileriosis are important hemoprotozoal diseases of sheep and goats in tropical and subtropical regions that lead to economic losses in these animals. PCR-restriction fragment length polymorphism (PCR-RFLP) is a reliable molecular diagnostic tool for discriminating Theileria or Babesia species in the same host. In this study, the DNA sequences of a ribosomal protein S8 (RPS8) gene from four species of piroplasms in China were used to develop a species-specific PCR-RFLP diagnostic tool. The sensitivity of the PCR assays was 0.1 pg DNA for B. motasi and 1 pg DNA for T. uilenbergi and 10 pg DNA for Babesia sp. Xinjiang-2005 and T. luwenshuni. The clear size difference of the PCR products allowed for a direct discrimination for B. motasi, Babesia sp. Xinjiang-2005 and ovine Theileria species (T. uilenbergi and T. luwenshuni), except that the mixed infection between T. uilenbergi and T. luwenshuni may be difficult to distinguish, simply after the electrophoretic separation of the amplification products. Further T. uilenbergi and T. luwenshuni diagnoses were made by digesting the PCR product with SacI. The established method could be applicable for the survey of parasite dynamics, and epidemiological studies as well as prevention and control of the disease.
绵羊泰勒虫病和巴贝斯虫病是热带和亚热带地区绵羊和山羊的重要血液原虫病，会给这些动物带来经济损失。聚合酶链反应-限制性片段长度多态性（PCR-RFLP）是一种可靠的分子诊断工具，可用于区分同一宿主中的泰勒虫属和巴贝斯虫属物种。在本研究中，使用来自中国的四种梨形虫的核糖体蛋白 S8（RPS8）基因的 DNA 序列开发了一种种特异性 PCR-RFLP 诊断工具。PCR 检测的灵敏度为 B. motasi 的 0.1 pg DNA、T. uilenbergi 的 1 pg DNA、Babesia sp. Xinjiang-2005 的 10 pg DNA 和 T. luwenshuni 的 10 pg DNA。PCR 产物的大小差异明显，可直接区分 B. motasi、新疆 2005 年巴贝斯虫和绵羊泰勒虫属（T. uilenbergi 和 T. luwenshuni）物种，只是 T. uilenbergi 和 T. luwenshuni 的混合感染可能难以区分，只需在电泳分离扩增产物后即可。进一步用 SacI 消化 PCR 产物可诊断 T. uilenbergi 和 T. luwenshuni。所建立的方法可适用于寄生虫动态调查、流行病学研究以及疾病的预防和控制。
