实时荧光PCR法检测包装饮用水中铜绿假单胞菌的研究  

Study on the Detection of Pseudomonas aeruginosa in Packaged Drinking Water by Real-Time Fluorescent PCR

作　　者：庞婕 刘杰 孔庆岩 秦森翰 董建锋 PANG Jie;LIU Jie;KONG Qingyan;QIN Senhan;DONG Jianfeng(Chengde Food and Drug Inspection and Testing Center,Chengde 067000,China;Innovation Center for Authentic Medicinal Materials Inspection and Testing in Yanshan District,Chengde 067000,China;Development Research Center of Hebei Provincial Administration for Market Regulation,Chengde 067000,China)

机构地区：[1]承德市食品药品检验检测中心,河北承德067000 [2]河北省燕山地区道地药材检验检测创新中心,河北承德067000 [3]河北省市场监督管理局发展研究中心,河北承德067000

出　　处：《食品安全导刊》2024年第6期69-71,75,共4页China Food Safety Magazine

基　　金：承德市科技计划项目(202303A167)。

摘　　要：近年来,包装饮用水中铜绿假单胞菌超标的现象时有发生,运用国家安全标准方法进行检验,过程烦琐,周期较长,不利于监管。然而,实时荧光PCR检验法具有灵敏、快速、准确等优点,被广泛应用于各种检验检测中。本文采用实时荧光PCR法,利用特异性引物和探针对目标基因进行扩增。然后通过实时荧光PCR法对其他5种细菌和梯度含量的铜绿假单胞菌菌悬液进行检测,验证方法的特异性和灵敏度。结果表明,只有铜绿假单胞菌的实时荧光PCR检测结果为阳性,其他细菌的检测结果为阴性,说明本方法对铜绿假单胞菌的特异性较好,方法的灵敏度为1×10^(3)CFU·mL^(-1)。同国家安全标准方法相比,实时荧光PCR法所用时间更短,明显提高了检测效率。In recent years,the phenomenon of excessive levels of Pseudomonas aeruginosa in packaged drinking water has occurred.The use of national safety standard methods for inspection is cumbersome and timeconsuming,which is not conducive to supervision.However,the real-time fluorescence PCR detection method has the advantages of sensitivity,speed,and accuracy,and is widely used in various testing methods.In this paper,realtime fluorescent PCR was used to amplify the target gene with specific primers and probes.Then,the other 5 kinds of bacteria and the gradient content of Pseudomonas aeruginosa suspensions were detected by real-time fluorescent PCR to verify the specificity and sensitivity of the method.The results showed that only Pseudomonas aeruginosa was positively detected by real-time fluorescent PCR,while the detection results of other bacteria were negative,indicating that the specificity of this method for Pseudomonas aeruginosa was good,and the sensitivity of this method was 1×10^(3) CFU·mL^(-1).Compared with the national safety standard method,the real-time fluorescent PCR method takes less time and significantly improves the detection efficiency.

关 键 词：铜绿假单胞菌 实时荧光PCR 包装饮用水 

分 类 号：R123.1[医药卫生—环境卫生学]