羊种布鲁氏菌中国优势流行株利福平抗性相关基因筛选及其抗性机制的初步研究

Brucella spp. is a facultative intracellular pathogen that can persistent infecting host cells, causing zoonotic diseases, and even affecting public health security and economic development. One of the brucellosis treatments recommended by World Health Organization was rifampicin. Studies have shown that the rpoB gene was associated with rifampicin resistance of Brucella spp.. The SNPs of this gene were results in the reduced sensitivity to rifampicin. At present, many rifampicin-resistant strains had been isolated from many countries and regions around the world, and these also had been reports in China in recent years. While some resistant strains had not detected SNPs of rpoB gene. Whether there were other rifampicin-resistance genes and mechanisms in the Brucella genome had not been clearly reported. Our team used the transposon random mutagenesis methods and whole genome sequencing of rifampicin-resistant strains to obtain 11 genes and regions related to rifampicin resistance in addition to rpoB gene before. In order to screen new rifampicin-metabolic genes and rapidly detect the rifampicin-resistance level of the Brucella isolated strains in China, this project used gene-recombination methods to detect the rifampicin-resistant strains. Meanwhile, the PCR detection method of rifampicin resistance of Brucella in China was established based on the screening genes. Using transcriptome sequencing and protein interaction technology to explore the mechanism of rifampicin resistance, this project could be lay the foundation for the prevention and control of Brucella spp. in China.

布鲁氏菌是一种兼性胞内寄生菌，引起人畜共患病，影响公共卫生安全和经济发展。利福平是世界卫生组织推荐的布鲁氏菌病治疗药物之一。研究表明，布鲁氏菌rpoB基因与利福平抗性相关。目前许多国家和地区已分离到利福平抗性菌株，但部分抗性菌株中未检测到rpoB基因变异，布鲁氏菌基因组中是否存在其他利福平抗性基因及机制尚无明确报道。为鉴定布鲁氏菌利福平抗性相关新基因，确定我国布鲁氏菌分离株利福平抗性水平及演化趋势，课题组前期开展了布鲁氏菌利福平抗性菌株RB51转座子随机突变以及布鲁氏菌利福平抗性人工诱变菌株全基因组测序分析，获得除rpoB基因外的11个利福平抗性候选基因。本项目拟利用同源重组技术鉴定利福平抗性相关新基因，建立一步法利福平抗性PCR检测方法，并利用转录组测序和蛋白互作技术筛选利福平抗性相关代谢通路，探讨利福平抗性的新机制，为我国布鲁氏菌分离株耐药性检测和利福平抗性菌株防控奠定基础。

布鲁氏菌是一种兼性胞内寄生菌，引起全球范围内流行的人畜共患的布鲁氏菌病，影响公共卫生安全和经济发展。利福平是世界卫生组织推荐的布鲁氏菌病治疗药物之一。研究表明，布鲁氏菌rpoB基因与利福平抗性相关。目前许多国家和地区已分离到利福平抗性菌株，但部分抗性菌株中未检测到rpoB基因变异，布鲁氏菌基因组中是否存在其他利福平抗性基因及机制尚无明确报道。本项目利用转录组和比较基因组分析并通过基因重组技术发现四型分泌系统及其涉及的代谢通路与低浓度利福平适应性耐受相关，确定我国布鲁氏菌分离株利福平抗性水平及演化趋势，为我国布鲁氏菌分离株耐药性检测和利福平抗性菌株防控奠定基础。