基于实时荧光聚合酶链式反应快速检测包装饮用水中铜绿假单胞菌

Rapid detection of Pseudomonas aeruginosa in packaged drinking water by real-time fluorescence polymerase chain reaction

中文关键词:  铜绿假单胞菌  实时荧光PCR  包装饮用水

英文关键词:Pseudomonas aeruginosa  real-time fluorescence polymerase chain reaction  packaged drinking water

中文摘要:

      目的 建立一种基于实时荧光聚合酶链式反应快速检测包装饮用水中铜绿假单胞菌的检测方法。方法 针对选择铜绿假单胞菌gyrB基因设计特异性引物和探针。250 mL水样经膜过滤后经过短时培养(36℃、4 h), 优化提取菌体DNA流程, 预冻干聚合酶链式反应预混液, 采用实时荧光聚合酶链式反应进行检测。结果 样品中铜绿假单胞菌量为≥1 CFU/250 mL时, 检测结果均呈阳性, 整个检测用时6 h左右。该方法检测结果与国家标准方法(GB 8538—2022)检测结果一致。结论 该方法特异性强、灵敏度高、用时短, 可为预包装饮用水中铜绿假单胞菌监管和生产企业质控提供更为快速、精准的技术手段。

英文摘要:

      Objective To establish a rapid detection method of Pseudomonas aeruginosa in packaged drinking water based on real-time fluorescence polymerase chain reaction. Methods Specific primer and probes were design for that selection of Pseudomonas aeruginosa gyrB gene. The 250 mL water sample was filtered through the membrane and subjected to short-term culture (36℃ and 4 h) to optimize the extraction process of bacterial DNA. The pre-mixed solution of polymerase chain reaction was lyophilized and detected by real-time fluorescent polymerase chain reaction. Results The test results were positive when the amount of Pseudomonas aeruginosa in the sample was≥1 CFU/250 mL, and the whole detection time was about 6 h. The test results by this method were consistent with those by the national standard method (GB 8538—2022). Conclusion This method has strong specificity, high sensitivity and short time, and can provide a more rapid and accurate technical means for the supervision of Pseudomonas aeruginosa in prepackaged drinking water and the quality control of production enterprises.