Yin Fangyuan, Liu Junlong, Liu Aihong, Li Youquan, Luo Jianxun, Guan Guiquan, 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 Science, Xujiaping 1, Lanzhou, Gansu 730046, PR China.
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Science, Xujiaping 1, Lanzhou, Gansu 730046, PR China.
Vet Parasitol. 2017 Apr 15;237:125-129. doi: 10.1016/j.vetpar.2017.02.019. Epub 2017 Feb 21.
Rapid and accurate diagnosis of Theileria annulata infection contributes to the formulation of strategies to eradicate this parasite. A simple and efficient diagnostic tool, recombinase polymerase amplification (RPA) combined with a lateral flow (LF) strip, was used in detection of Theileria and compared to other methods that require expensive instruments and skilled personnel. Herein, we established and optimized an LF-RPA method to detect the cytochrome b gene of T. annulata mitochondrial DNA from experimentally infected and field-collected blood samples. This method has many unparalleled characteristics, including that it is rapid (clear detection in 5min at constant temperature), sensitive (the limitation of detection is at least 2pg genomic DNA), and specific (no cross-reaction with other piroplasms that infect cattle). The LF-RPA assay was evaluated via testing 17 field blood samples and comparing the results of that of a PCR, showing 100% agreement, which demonstrates the ability of the LF-RPA assay to detect T. annulata infections in small number of samples (n=17). Taken together, the results indicate that this method could be used as an ideal diagnostic tool for detecting T. annulata in endemic regions with limited to fewer and local resources and could also be a potential technique for the surveillance and control of blood protozoa.
环形泰勒虫感染的快速准确诊断有助于制定根除这种寄生虫的策略。一种简单有效的诊断工具，即重组酶聚合酶扩增（RPA）结合侧向流动（LF）试纸条，被用于检测泰勒虫，并与其他需要昂贵仪器和专业人员的方法进行了比较。在此，我们建立并优化了一种LF-RPA方法，用于从实验感染和现场采集的血液样本中检测环形泰勒虫线粒体DNA的细胞色素b基因。该方法具有许多无与伦比的特点，包括快速（恒温5分钟即可清晰检测）、灵敏（检测限至少为2pg基因组DNA）和特异（与其他感染牛的梨形虫无交叉反应）。通过检测17份现场血液样本并将LF-RPA检测结果与PCR结果进行比较，对LF-RPA检测进行了评估，结果显示二者100%一致，这证明了LF-RPA检测在少量样本（n=17）中检测环形泰勒虫感染的能力。综上所述，结果表明该方法可作为在资源有限的地方性流行地区检测环形泰勒虫的理想诊断工具，也可能是监测和控制血液原虫的一种潜在技术。
