华南理工大学学报(自然科学版) ›› 2004, Vol. 32 ›› Issue (5): 51-55.

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菌落PCR方法的建立及其与常规PCR方法的比较

徐丽 蔡俊鹏   

  1. 华南理工大学 食品与生物工程学院, 广东 广州 510640
  • 收稿日期:2003-07-12 出版日期:2004-05-20 发布日期:2015-09-08
  • 通信作者: 徐丽(1978-), 女, 主要从事食品生物技术方面的研究. E-mail:nancy_xu0775@163.com
  • 作者简介:徐丽(1978-), 女, 主要从事食品生物技术方面的研究.
  • 基金资助:
    国家自然科学基金资助项目(30070591,40176036)

Establishment of Colony PCR Method and Its Comparison with Conventional PCR Method

Xu Li  Cai Jun-peng   

  1. Establishment of Colony PCR Method and Its Comparison with Conventional PCR Method
  • Received:2003-07-12 Online:2004-05-20 Published:2015-09-08
  • Contact: 徐丽(1978-), 女, 主要从事食品生物技术方面的研究. E-mail:nancy_xu0775@163.com
  • About author:徐丽(1978-), 女, 主要从事食品生物技术方面的研究.

摘要: 为建立一种快速 、经济、敏感的 PCR 检测方法 ,以直接检测出环境 、海产品等中的靶基因 ,通过PCR扩增已知携带 tlh 和 tdh基因的副溶血弧菌标准菌株, 并和常规 PCR方法比较 ,初步建立了菌落 PCR技术 ;再以副溶血弧菌标准菌株为阳性对照, 以本实验室自海洋环境中分离得到的数株野生菌株为检测对象, 采用新建立的菌落 PCR 以及常规 PCR方法对它们进行扩增比较, 进一步证明了菌落 PCR 技术检测携带相关基因菌株的可靠性.通过比较,发现菌落 PCR 和常规 PCR的结果相当一致, 两种方法均能扩增出阳性对照标准菌株中 tlh 和 tdh 基因.

关键词: 菌落 PCR, 常规 PCR, tdh 基因 , tlh 基因

Abstract: To establish a quick, sensitive and economical PCRmethod for directly detecting tlh and adh genes-carrying Vibrio parahaemolyticus in environment and seafood, a colony PCR method was developed and its resultswere compared with those of the conventional PCRmethod, thus establishing a colony PCRmethod.By using the Vibrioparahaemolytic-us strains as positive controls, several marine bacteria strains preserved in the lab were amplified through the colony PCR method and the conventional PCR method , by which the reliability of colony PCR method for the corresponding gene de-tection was proved.The comparison shows that the results obtained by colony PCR method are in great agreement with those obtained by the conventional PCRmethod, and that the tlh and tdh genes in positive controls can be amplified by these two PCR methods.

Key words: colony PCR, conventional PCR, tdh gene, tlh gene

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