Brucellosis is a worldwide re-emerging zoonosis. It has an economic impact due to abortion and loss of fertility in livestock. In this study, Real-time recombinase polymerase amplification (RT-RPA-BP26) targeting Brucella spp. bp26 gene and Lateral flow dipstick (LFD-RPA-IS711) combined with SYBR- Green recombinase polymerase amplification (RPA) targeting insertion sequence IS711 region of Brucella spp. bp26 gene, was developed to detect Brucella spp. from different sample types in domestic animals. The sensitivity and specificity of the two developed RPAs were compared with real-time PCR, PCR, and Rose Bengal Plate Test (RBPT). The analytical sensitivity and detection limit of Real-time RPA and LFD RPA were four and six copies per reaction respectively. The detection of six colony forming units (CFU) of the bacteria-bearing construct with the target sequence was within 20 min at 40 °C for Real-time RPA and 37 °C for LFD RPA. The LFD RPA could work at temperatures between 30 and 35 °C and could be completed within 10-30 min. No significant differences were observed when comparing the results from Real-time RPA and LFD RPA to Real-time PCR and PCR. Both methods showed no cross reactivity with Chlamydia abortus, Toxoplasma gondii, Salmonella typhimurium, and Escherichia coli. In conclusion, RPA is a useful and convenient field and point of care test for brucellosis.

中文翻译：

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建立用于检测布鲁氏菌属的重组酶聚合酶扩增（RPA）分析方法。感染。

布鲁氏菌病是一种世界范围内正在重新出现的人畜共患病。由于牲畜的流产和丧失生育能力，对经济产生影响。在这项研究中，针对布鲁氏菌属的实时重组酶聚合酶扩增（RT-RPA-BP26）。bp26基因和侧向量油尺（LFD-RPA-IS711）与SYBR- Green重组酶聚合酶扩增（RPA）结合，靶向布鲁氏菌属的插入序列IS711区。bp26基因被开发用于检测布鲁氏菌属。来自家畜的不同样本类型。将两个已开发的RPA的敏感性和特异性与实时PCR，PCR和玫瑰孟加拉平板试验（RBPT）进行了比较。实时RPA和LFD RPA的分析灵敏度和检出限分别为每个反应4个和6个拷贝。对于带有目标序列的带有细菌的构建体，六个菌落形成单位（CFU）的检测在实时RTA的40°C和LFD RPA的37°C下于20分钟内完成。LFD RPA可以在30到35°C的温度下工作，并且可以在10到30分钟内完成。将实时RPA和LFD RPA的结果与实时PCR和PCR进行比较时，没有观察到显着差异。两种方法均显示与流产衣原体，弓形体，鼠伤寒沙门氏菌和大肠杆菌无交叉反应。总之，RPA是布鲁氏菌病有用且方便的现场检查点。将实时RPA和LFD RPA的结果与实时PCR和PCR进行比较时，没有观察到显着差异。两种方法均显示与流产衣原体，弓形体，鼠伤寒沙门氏菌和大肠杆菌无交叉反应。总之，RPA是布鲁氏菌病有用且方便的现场检查点。将实时RPA和LFD RPA的结果与实时PCR和PCR进行比较时，没有观察到显着差异。两种方法均显示与流产衣原体，弓形体，鼠伤寒沙门氏菌和大肠杆菌无交叉反应。总之，RPA是布鲁氏菌病有用且方便的现场检查点。