食品科学 ›› 2010, Vol. 31 ›› Issue (16): 270-273.doi: 10.7506/spkx1002-6630-201016059

• 分析检测 • 上一篇    下一篇

环介导等温扩增技术检测肉及肉制品中沙门氏菌

王 羽,王贞强,张 伟*   

  1. 河北农业大学食品科技学院
  • 收稿日期:2010-06-22 出版日期:2010-08-25 发布日期:2010-12-29
  • 通讯作者: 张 伟* E-mail:zhangwei631126@163.com
  • 基金资助:

    河北省自然科学基金项目(C2008000216)

Loop-mediated Isothermal Amplification Assay for the Detection of Salmonella in Meat and Meat Products

WANG Yu,WANG Zhen-qiang,ZHANG Wei*   

  1. (College of Food Science and Technology, Agricultural University of Hebei, Baoding 071000, China)
  • Received:2010-06-22 Online:2010-08-25 Published:2010-12-29
  • Contact: ZHANG Wei* E-mail:zhangwei631126@163.com

摘要:

建立用环介导等温扩增技术(loop-mediated isothermal amplification,LAMP)检测肉及肉制品中沙门氏菌的方法。利用靶基因的6 个区段设计4 条引物,应用具有链置换活性的Bst DNA 聚合酶,能够在恒温(65℃左右)条件下特异、高效、快速地扩增待测物的DNA。针对沙门氏菌的特异基因invA 基因设计两对LAMP 引物,建立LAMP 检测沙门氏菌的新方法,并对肉及肉制品中的沙门氏菌进行检测。结果表明:建立LAMP 技术检测肉及肉制品中沙门氏菌的方法,该方法能够特异检测沙门氏菌,且灵敏性可达10-9CFU/mL,对48 份样品检测,该方法检出率为91.6%、敏感性100%、特异性70.0%、符合率为93.8%、检出时间1.5h。

关键词: 环介导等温扩增, 检测, 沙门氏菌, 肉及肉制品

Abstract:

A novel nucleic acid amplification method, termed loop-mediated isothermal amplification (LAMP), which amplifies DNA with high specificity, efficiency, and rapidity under isothermal conditions, may be a valuable tool for the rapid detection of infectious diseases. In this study, two pairs of gene-specific primers for the conservative invA gene were designed to establish a LAMP assay for detecting Salmonella. The assay method was applied to detect meat of different animal species, and proved highly sensitive and specific and time saving.

Key words: loop-mediated isothermal amplification, detection, Salmonella, meat and meat products

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