Establishment of a recombinase polymerase amplification (RPA) assay for the detection of Brucella spp. Infection

文献类型： 外文期刊

第一作者： Gumaa, M. M.

作者： Gumaa, M. M.;Cao, Xiaoan;Li, Zhaocai;Lou, Zhongzi;Zhang, Nianzhang;Zhang, Zhijun;Zhou, Jizhang;Fu, Baoquan

作者机构：

关键词： Brucellosis; Recombinase polymerase amplification; PCR; Real-time PCR; bp26 gene lateral-flow dipstick; SYBR green I; IS711

期刊名称：MOLECULAR AND CELLULAR PROBES （ 影响因子：2.365； 五年影响因子：2.386 ）

ISSN： 0890-8508

年卷期： 2019 年 47 卷

页码：

收录情况： SCI

摘要： 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 degrees C for Real-time RPA and 37 degrees C for LFD RPA. The LFD RPA could work at temperatures between 30 and 35 degrees 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.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序