Brucella, the etiological agent of brucellosis, is an important zoonosis pathogen worldwide. Brucella infects humans and various domestic and wild animals, and represents a great threat to public health and animal husbandry. In the present study, we developed a real-time recombinase polymerase amplification (RPA) assay for the detection of Brucella. The assay targeted the bcsp31 gene of Brucella, and an RPA exo probe and a pair of primers were selected for assay validation. RPA sensitivity and specificity were evaluated using plasmid standards, Brucella representative strains, and non-Brucella strains. The RPA assay achieved a detection limit of 17 molecules in 95% of cases based on probit analysis, and could successfully distinguish 18 representative Brucella strains (B. abortus biovars 1, 2, 3, 4, 5, 6, 7 and 9, B. melitensis biovars 1, 2 and 3, B. suis biovars 1, 2, 3 and 4, B. canis, B. neotomae and B. ovis), and four Brucella vaccine strains (A19, S19, S2 and M5). A total of 52 Brucella field strains were detected by real-time PCR and RPA in parallel, and compared with real-time PCR, the sensitivity of the RPA assay was 94% (49/52). Thus, this RPA assay may be a rapid, sensitive, and specific tool for the prevention and control of Brucellosis.

中文翻译：

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一种使用实时重组酶聚合酶扩增测定法检测布鲁氏菌的新方法。

布鲁氏菌病是布鲁氏菌病的病原体，是全世界重要的人畜共患病病原体。布鲁氏菌感染人类以及各种家畜和野生动物，对公共卫生和畜牧业构成巨大威胁。在本研究中，我们开发了一种实时重组酶聚合酶扩增（RPA）检测方法，用于检测布鲁氏菌。该测定法针对布鲁氏菌的bcsp31基因，并选择RPA exo探针和一对引物进行测定法验证。使用质粒标准品，布鲁氏菌代表性菌株和非布鲁氏菌菌株评估RPA敏感性和特异性。根据概率分析，RPA分析在95％的病例中达到了17个分子的检出限，并且可以成功地区分18个代表性的布鲁氏菌菌株（B.abortus biovars 1，2，3，4，5，6，7和9，B 。melitensis biovars 1、2和3，猪双歧杆菌，1、2、3和4，犬双歧杆菌，新芽孢杆菌和卵形芽孢杆菌，以及四个布鲁氏菌疫苗株（A19，S19，S2和M5）。实时PCR和RPA并行检测到52株布鲁氏菌田间菌株，与实时PCR相比，RPA测定的灵敏度为94％（49/52）。因此，该RPA测定法可能是用于预防和控制布鲁氏菌病的快速，灵敏且特定的工具。