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 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 Sciences, Xujiaping 1, Lanzhou, Gansu, 730046, PR China.
Ticks Tick Borne Dis. 2018 May;9(4):1002-1005. doi: 10.1016/j.ttbdis.2018.03.028. Epub 2018 Mar 28.
Theileriosis is a disease of domesticated animals in tropical and subtropical countries and causes significant reductions in livestock productivity. The arid region of Punjab in Pakistan is notorious for the presence of the vector tick (Acari: Ixodidae) and tick-borne diseases, such as theileriosis and babesiosis. The distribution of Theileria annulata and T. orientalis in the Chakwal district of Punjab was determined by developing a multiplex recombinase polymerase amplification (RPA) assay as a scientific basis for formulating control strategies for bovine theileriosis. Specificity was evaluated using DNA from related piroplasm species, while analytical sensitivity was calculated using a long fragment of the enolase gene. A total of 188 blood samples were collected on FTA cards (Whatman) from tick-infested asymptomatic breeds of cattle (Bos indicus, Bos taurus, and Bos indicus × Bos taurus) in the study district. Finally, infections with of T. annulata and T. orientalis were detected using the multiplex RPA and compared with the conventional multiplex polymerase chain reaction (PCR). The multiplex RPA specifically amplified 282-bp and 229-bp fragments of the enolase gene from T. annulata and T. orientalis and had no cross-reaction with other piroplasm species. It was determined that 45 (23.9%) and 5 (2.6%) out of 188 blood samples were positive for T. annulata and T. orientalis, respectively, when examined using RPA. Multiplex PCR detection indicated that 32 (17.0%) and 3 (1.6%) blood samples were positive for T. annulata and T. orientalis, respectively. In the present study, a specific RPA method was developed for simultaneous differentiation and detection of T. annulata and T. orientalis infections and used for the first time for the detection of the two bovine Theileria infections.
泰勒虫病是热带和亚热带国家家养动物的一种疾病，会导致牲畜生产力显著下降。巴基斯坦旁遮普省的干旱地区以存在媒介蜱（蜱螨目：硬蜱科）和蜱传疾病而闻名，如泰勒虫病和巴贝斯虫病。通过开发多重重组酶聚合扩增（RPA）检测方法，确定了旁遮普省查克瓦尔地区的环形泰勒虫和东方泰勒虫的分布，这为制定牛泰勒虫病的控制策略提供了科学依据。使用相关梨形虫种的 DNA 评估了特异性，而通过烯醇酶基因的长片段计算了分析灵敏度。总共从受感染无症状的牛品种（印度野牛、黄牛和印度野牛×黄牛）收集了 188 份血液样本，这些样本是在研究地区用 FTA 卡（沃特曼）采集的。最后，使用多重 RPA 检测到 T. annulata 和 T. orientalis 的感染，并与传统的多重聚合酶链反应（PCR）进行了比较。多重 RPA 特异性地扩增了 T. annulata 和 T. orientalis 烯醇酶基因的 282-bp 和 229-bp 片段，与其他梨形虫种没有交叉反应。通过 RPA 检测，188 份血液样本中有 45 份（23.9%）和 5 份（2.6%）分别为 T. annulata 和 T. orientalis 阳性。多重 PCR 检测表明，32 份（17.0%）和 3 份（1.6%）血液样本分别为 T. annulata 和 T. orientalis 阳性。在本研究中，开发了一种用于同时区分和检测 T. annulata 和 T. orientalis 感染的特异性 RPA 方法，并首次用于检测这两种牛泰勒虫感染。
