食品科学

• 分析检测 • 上一篇    下一篇

食品过敏原澳洲坚果环介导等温扩增检测方法的建立与应用

刘 津1,张 隽1,李 婷1,张 璜2,高东微1,李志勇1,*   

  1. 1.广东检验检疫技术中心,广东 广州 510623;2.广州迪澳生物科技有限公司,广东 广州 510663
  • 出版日期:2014-11-25 发布日期:2014-12-09
  • 基金资助:

    出入境检验检疫行业标准制定计划项目(2012B141)

Construction and Application of Loop-Mediated Isothermal Amplification Method for Detection of Macadamia Nut as a Food Allergen

LIU Jin1, ZHANG Jun1, LI Ting1, ZHANG Huang2, GAO Dong-wei1, LI Zhi-yong1,*   

  1. 1. Guangdong Inspection and Quarantine Technology Center, Guangzhou 510623, China;
    2. Guangzhou Di Ao Bio-Technology Co. Ltd., Guangzhou 510663, China
  • Online:2014-11-25 Published:2014-12-09

摘要:

根据澳洲坚果豌豆蛋白AMP2基因序列,利用设计软件Primer Explorer Version 4设计并筛选了食品过敏原澳洲坚果的环介导等温扩增引物,对反应体系和反应条件进行优化,建立澳洲坚果的环介导等温扩增检测方法,结果判断可采用实时荧光法和荧光染料终点显色法。对该方法进行了特异性、灵敏度、稳定性评价,结果显示:该方法能够特异性、灵敏、稳定地检测食品中的澳洲坚果成分,检测低限为0.5%。此外,对7 种市售食品样品的检测结果表明,该方法与食品标签标示的过敏原成分结果吻合率为100%,假阳性率和假阴性率均为0,在市售食品的过敏原成分检测上较商业化快速检测试纸条更加稳定可靠。

关键词: 食品过敏原, 澳洲坚果, 环介导等温扩增, 检测方法

Abstract:

According to the Macadamia integrifolia vicilin precursor AMP2 gene sequences, sets of loop-mediated
isothermal amplification (LAMP) primers were designed and screened using the design software Primer Explorer Version
4, then the relevant LAMP method was established for the detection of macadamia nuts as a food allergen by measuring the
results using real-time fluorescence or using a fluorescent dye in the reaction end. An evaluation of this method indicated
the satisfied specificity, sensitivity and stability in the identification of macadamia nuts from foods with a LOD of 0.5%. In
addition, the detection of 7 marketed food samples using this method showed 100% consistency with the labeled information
of allergen ingredients with zero false positive and negative rates, and thus, proved its better performances than commercial
rapid test strips.

Key words: food allergen, macadamia nut, loop-mediated isothermal amplification (LAMP), detection method

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