食品科学 ›› 2009, Vol. 30 ›› Issue (6): 139-142.doi: 10.7506/spkx1002-6630-200906029

• 分析检测 • 上一篇    下一篇

变性高效液相色谱检测乳制品和化妆品中绿脓杆菌的研究

曹际娟1,郑秋月1.2孙哲平1,王秋艳1,裴轶君1,赵昕1   

  1. 1. 辽宁出入境检验检疫局
    2.沈阳农业大学
  • 收稿日期:2008-05-19 修回日期:2008-06-30 出版日期:2009-03-15 发布日期:2010-12-29
  • 通讯作者: 曹际娟 E-mail:cjj0909@163.com
  • 基金资助:

    “十一五”国家科技支撑计划项目(2006BAK02A13)

Denaturing High-Performance Liquid Chromatography Coupled with Polymerase Chain Reaction for Detecting seudomonas aeruginosa in Dairy and Cosmetic Products

CAO Ji-juan1,ZENG Qiu-yue1,2,SUN Zhe-ping1,WANG Qiu-yan1,PEI Yi-jun1,ZHAO Xin1   

  1. (1. Liaoning Entry-Exit Inspection and Quarantine Bureau, Dalian 116001, China;
    2. Shenyang Agricultural University, Shenyang 110161, China)
  • Received:2008-05-19 Revised:2008-06-30 Online:2009-03-15 Published:2010-12-29
  • Contact: CAO Ji-juan1 E-mail:cjj0909@163.com

摘要:

目的:应用PCR 结合变性高效液相色谱技术实现绿脓杆菌的快速鉴定。方法:利用所报道的绿脓杆菌外毒素A(ETA)基因设计引物和探针,并对该方法进行特异性、灵敏度、精密度等方面的考察。结果:用该方法未检测到绿脓杆菌的近源种及其他细菌的阳性吸收峰,检测灵敏度可达到100CFU/ml。结论:用PCR 结合变性高效液相色谱检测绿脓杆菌不仅特异性好、灵敏度高,且快速简便。

关键词: DHPLC, 脓杆菌, 外毒素A基因, 检测

Abstract:

Objective: To rapidly identify Pseudomonas aeruginosa using denaturing high-performance liquid chromatography DHPLC) coupled with polymerase chain reaction (PCR). Methods: The primers and probes for specific PCR amplification ere designed based on the reported sequence of exotoxin type A (ETA)gene of P.aeruginosa, and then the specificity, sensitivity nd stability of this method were analyzed. Results: In analysis of Pseudomonas aeruginosa and other species of Pseudomonas y this method, only P.aeruginosa can be detected, and the sensitivity is 100 CFU/ml. Conclusion: The PCR-DHPLC method s rapid, specific, sensitive and relatively simple for the identification of P.aeruginosa.

Key words: DHPLC, seudomonas aeruginosa, exotoxin type A gene, detection

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