实时荧光环介导等温扩增检测铜绿假单胞菌

Detection of Pseudomonas aeruginosa by real-time loop-mediated isothermal amplification

中文关键词:  实时荧光环介导等温扩增  铜绿假单胞菌  矿泉水

英文关键词:real-time loop-mediated isothermal amplification  Pseudomonas aeruginosa  mineral water

中文摘要:

      目的 建立一种实时荧光与环介导等温扩增(loop-mediated isothermal amplification, LAMP)相结合的铜绿假单胞菌检测方法。方法 利用铜绿假单胞菌的外毒素A基因(Pseudomonas aeruginosa exotoxin A, PEA)序列, 设计LAMP引物, 对该方法的特异性、灵敏度及其在人工污染矿泉水中的检出限进行评价。结果 该方法与其他菌株无交叉反应, 特异性强, 对纯菌液和人工污染矿泉水的检出限均可达到5.2 CFU/mL。结论 该方法快速、灵敏、操作简便, 为水样中铜绿假单胞菌的检测提供了一种有效的手段。

英文摘要:

      Objective To establish a detection method for Pseudomonas aeruginosa by real-time loop-mediated isothermal amplification (LAMP). Methods The LAMP primers were designed for the sequence of Pseudomonas aeruginosa exotoxin A (PEA). The specificity, sensitivity and detection limit of artificially polluted mineral water were measured to evaluate applicability of the LAMP assay. Results This method had no cross-reaction with other strains, which had strong specificity to P. aeruginosa. The detection limit in pure cultures and artificially-contaminated mineral water samples without enrichment was 5.2 CFU/mL. Conclusion This method is rapid, sensitive and easy to operate, which is suitable for detection of P. aeruginosa in water samples.