LAMP法检测食品中开心果过敏原成分

doi:10.7506/spkx1002-6630-201322026

食品科学 ›› 2013, Vol. 34 ›› Issue (22): 128-132.doi: 10.7506/spkx1002-6630-201322026

LAMP法检测食品中开心果过敏原成分

刘昊，黄文胜，邓婷婷，傅凯，鞠兴荣，陈颖

1.南京财经大学食品科学与工程学院，江苏南京 210003；2.中国检验检疫科学研究院农产品安全研究中心，北京 100123

收稿日期:2013-01-17 修回日期:2013-10-28 出版日期:2013-11-25 发布日期:2013-12-05
通讯作者: 陈颖 E-mail:chenyingcaiq@163.com
基金资助:
国家自然科学基金项目;国家“十二五”科技支撑计划项目

Detection of Pistachio Allergen in Foods by Loop-Mediated Isothermal Amplification Method

LIU Hao，HUANG Wen-sheng，DENG Ting-ting，FU Kai，JU Xing-rong，CHEN Ying

1. College of Food Science and Engineering, Nanjing University of Finance and Economics, Nanjing 210003, China；
2. Agro-product Safety Research Center, Chinese Academy of Inspection and Quarantine, Beijing 100123, China

Received:2013-01-17 Revised:2013-10-28 Online:2013-11-25 Published:2013-12-05
Contact: ying chen E-mail:chenyingcaiq@163.com

摘要/Abstract

摘要：

为建立食品中开心果过敏原成分的环介导等温扩增(LAMP)检测方法，根据GenBank中公布的开心果过敏原2S种子贮存蛋白基因序列设计合成LAMP引物，筛选最佳引物并对其内引物浓度进行优化。以开心果、腰果、胡桃等坚果类以及大米、玉米、大豆等常见农产品验证该方法的特异性，将开心果DNA进行梯度稀释以验证方法的检测低限，并以小麦为基质配制模拟添加样品测试方法的相对灵敏度。结果表明：该方法能够特异性检测食品中的开心果成分，对开心果基因组DNA的检测限为0.2ng，食品中开心果成分相对检测灵敏度低于0.1%(m/m)，通过在反应终产物中加入SYBR GreenⅠ，可用肉眼判断检测结果，LAMP检测时间不超过60min。该方法快速准确，不依赖复杂昂贵的检测仪器，可应用于过敏原开心果成分的快速检测。

关键词: 过敏原, 开心果, 环介导等温扩增, 检测

Abstract:

Loop-mediated isothermal amplification (LAMP) was applied for the rapid detection of pistachio allergen in
foods. According to the sequence of Pistacia vera (Pis v1) allergen 2S albumin, a set of primers was designed. Various
LAMP parameters including reaction temperature and inner primer concentration were optimized and finally the optimal
LAMP assay for the detection of pistachio allergen in foods was established. The specificity, sensitivity and robustness of
the developed method were tested. The results indicated that no cross reaction was observed with other common tree nuts
such as pistachio, cashew, walnut or crops such as soybean, maize, wheat and rice. The absolute limit of detection for pistachio
DNA was 0.2 ng/reaction and the relative limit of detection was 0.1% (m/m) for simulated samples made by mixed
pistachio with wheat as the matrix. Since the LAMP assay could be completed within 60 minutes, only a regular water bath
was required, and the results could be observed by the naked eye according to the color change produced by SYBR GreenⅠ.
This method allows easy, rapid and accurate detection of pistachio allergen in foods and can be especially applicable in a
resource-limited situation.

Key words: allergen, pistachio, loop-mediated isothermal amplification (LAMP), detection

中图分类号:

TS207.3

刘昊，黄文胜，邓婷婷，傅凯，鞠兴荣，陈颖. LAMP法检测食品中开心果过敏原成分[J]. 食品科学, 2013, 34(22): 128-132.

LIU Hao，HUANG Wen-sheng，DENG Ting-ting，FU Kai，JU Xing-rong，CHEN Ying. Detection of Pistachio Allergen in Foods by Loop-Mediated Isothermal Amplification Method[J]. FOOD SCIENCE, 2013, 34(22): 128-132.

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LAMP法检测食品中开心果过敏原成分

Detection of Pistachio Allergen in Foods by Loop-Mediated Isothermal Amplification Method

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