食品科学

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酶连接探针杂交芯片特异性检测转基因水稻品系

张明哲1,陈吴健1,张晓峰2,陈笑梅2,陈 曦1,吴 蓉3   

  1. 1.浙江省检验检疫科学技术研究院,浙江 杭州 310016;2.浙江出入境检验检疫局,浙江 杭州 310016;
    3.杭州出入境检验检疫局,浙江 杭州 310012
  • 出版日期:2014-04-25 发布日期:2014-05-13
  • 基金资助:

    浙江省科技厅重大科技专项重点农业项目(2011C12023);国家质检总局科技计划项目(2011IK249);
    浙江省科技厅公益性项目(2011C22065);浙江出入境检验检疫局科技计划项目(ZK200958)

Event-specific Detection of Genetically Modified Rice Using Enzyme-Linked Probe Hybridization Chip

ZHANG Ming-zhe1, CHEN Wu-jian1, ZHANG Xiao-feng2, CHEN Xiao-mei2, CHEN Xi1, WU Rong3   

  1. 1. Zhejiang Academy of Science and Technology for Inspection and Quarantine, Hangzhou 310016, China;
    2. Zhejiang Entry-Exit Inspection and Quarantine Bureau, Hangzhou 310016, China;
    3. Hangzhou Entry-Exit Inspection and Quarantine Bureau, Hangzhou 310012, China
  • Online:2014-04-25 Published:2014-05-13

摘要:

酶连接探针杂交芯片是一种基于连接酶催化的探针杂交芯片技术,其原理是采用硫代引物保护法,利用 Lambda DNA外切酶将多重聚合酶链式反应产物切割成单链。在氨基修饰的芯片上固定5’端探针,并与制备的产物 单链进行杂交,加入荧光修饰的第2个探针与之前的探针进行酶连接反应,从而实现高特异性、高通量检测。结果 表明:酶连接探针杂交芯片技术检测4 种转基因水稻品系特异性好,灵敏度可达0.1%(m/m),可以应用于进出口 检验检疫、食品安全监测等领域。

关键词: 酶连接探针杂交芯片, 转基因水稻, 品系检测

Abstract:

The enzyme-linked probe hybridization chip is a method based on ligase-hybridizing probe chip technology with
the principle of using thio-primers for protection, and using Lambda DNA exonuclease to cut multiple PCR amplicons into
single products. 5’-End probe chain is fixed in the amino-modified chip, and hybridized with the single-chain product by
adding the fluorescent-modified probe for the enzyme-linked reactions with the anterior probe in order to achieve highly
specific, parallel and high-throughput detection. In this study, the enzyme-linked probe hybridization chip could effectively
detect genetically modified rice with high specificity and the sensitivity limit was 0.1%. This chip can be used by the entryexit
inspection and quarantine authorities and be applied for food safety testing.

Key words: enzyme-linked probe hybridization chip, genetically modified rice, event-specific detection

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