华南理工大学学报(自然科学版) ›› 2012, Vol. 40 ›› Issue (5): 149-154.

• 食品科学与技术 • 上一篇    

禽肉中弯曲菌的分离、PFGE 和DGGE 分型

孟赫诚1 毕水莲2 闫鹤1,3 1†   

  1. 1.华南理工大学 轻工与食品学院,广东 广州 510640; 2.广东药学院 食品科学学院,广东 中山 528435;3.辽宁省食品安全重点实验室,辽宁 锦州 121013
  • 收稿日期:2011-12-20 修回日期:2012-03-01 出版日期:2012-05-25 发布日期:2012-03-31
  • 通信作者: 石磊(1961-) ,男,博士,教授,主要从事病原微生物快速诊断和耐药机制研究. E-mail: leishi@scut.edu.cn E-mail:arsenal0001@ hotmail.com
  • 作者简介:孟赫诚(1984-) ,男,博士,主要从事微生物快速诊断和分子耐药研究.
  • 基金资助:

    广东省自然科学基金资助项目( 10451064101005159) ; 华南理工大学中央高校基本科研业务费专项资金资助项目( 2012ZZ0083) ; 辽宁省食品安全重点实验室开放课题资助项目( LNSAKF2011002)

Isolation of Campylobacter Strains in Poultry Products and Genotyping Identification of Strains by Means of PFGE and DGGE

Meng He-chengBi Shui-lian Yan He1,3  Shi Lei1   

  1. 1. School of Light Industry and Food Sciences,South China University of Technology,Guangzhou 510640,Guangdong,China; 2. School of Food Science,Guangdong Pharmaceutical University,Zhongshan 528435,Guangdong,China; 3. Food Safety Key Lab of Liaoning Province,Jinzhou 121013,Liaoning,China
  • Received:2011-12-20 Revised:2012-03-01 Online:2012-05-25 Published:2012-03-31
  • Contact: 石磊(1961-) ,男,博士,教授,主要从事病原微生物快速诊断和耐药机制研究. E-mail: leishi@scut.edu.cn E-mail:arsenal0001@ hotmail.com
  • About author:孟赫诚(1984-) ,男,博士,主要从事微生物快速诊断和分子耐药研究.
  • Supported by:

    广东省自然科学基金资助项目( 10451064101005159) ; 华南理工大学中央高校基本科研业务费专项资金资助项目( 2012ZZ0083) ; 辽宁省食品安全重点实验室开放课题资助项目( LNSAKF2011002)

摘要: 随机选取某肉菜市场8 份鸡肉和鸭肉样品中的弯曲菌( Campylobacter) ,分别采用Bolton 和Preston 肉汤增菌,然后以Skirrow、mCCDA 选择分离培养和膜过滤的方法同时分离,运用脉冲场凝胶电泳( PFGE) 和变性梯度凝胶电泳( DGGE) 对分离出的菌株进行分子水平的分型和溯源.结果表明: 共有6 份样品污染了空肠弯曲菌( C. jejuni) 或结肠弯曲菌( C. coli) ; Preston 肉汤增菌、mCCDA 选择分离培养法的分离效果最好; 4 份样品被同一种C. coli 污染,2 份样品分别污染了2 种C. jejuni 和3 种C. coli; 与PFGE 相比,DGGE分型方法的灵敏度略低,但更快速、成本更低,且稳定、准确,可用于食品中弯曲菌的快速分型.

关键词: 弯曲菌, 分离, 分型, 脉冲场凝胶电泳, 变性梯度凝胶电泳

Abstract:

Eight poultry samples including chicken and duck were collected in a certain fresh market and were enriched in Bolton and Preston broths,followed by the inoculation on selective agar media ( Skirrow and mCCDA) and by the membrane filtration. Then,the molecular types of the isolated Campylobacter strains were analyzed by means of pulse field gel electrophoresis ( PFGE) and denaturing gradient gel electrophoresis ( DGGE) ,and C. jejuni or C. coli were isolated from six samples. The results show that the combination of Preston broth and mCCDA media is the optimal isolation method,that there are four samples contaminated by the same C. coli isolate and two samples contaminated respectively by 2 species of C. jejuni and 3 species of C. coli,and that,though DGGE genotyping is not as sensitive as PFGE genotyping,it is more suitable for the rapid genotyping of Campylobacter in food borne because it is of high speed,low cost as well as stable and accurate operation.

Key words: Campylobacter, isolation, genotyping identification, pulse field gel electrophoresis, denaturing gradient gel electrophoresis