1种高特异性的酶连接探针杂交芯片及其在转基因检测中的应用  

Specific Detection of Genetic Modified Organism(GMO) Using Enzyme-linked Probe Hybridization Chip

作　　者：张明哲 张晓峰 陈笑梅 陈曦 吴姗 陈吴健 

机构地区：[1]浙江省检验检疫科学技术研究院,杭州310016

出　　处：《中国食品学报》2014年第10期223-227,共5页Journal of Chinese Institute Of Food Science and Technology

基　　金：浙江省科技厅重大专项(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)

摘　　要：酶连接探针杂交芯片是1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PCR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,从而实现高度特异性、高通量检测。特异性和灵敏度试验表明,酶连接探针杂交芯片能有效消除假阳性信号,转基因检测的灵敏度达到0.1%,可应用于进出口检验检疫、食品安全监测等领域。The enzyme-linked probe hybridization chip is a method based on ligase-hybridizing probe chip technology, with the principle of using the thio-primers for protection, and using lambda DNA exonuclease to cut multiple PCR products into single chains. 5′-end probe was fixed in the amino-modified chip, and hybridized with the single-chain product, 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. According to the specificity and sensitivity experiments,the enzyme-linked probe hybridization chip could effectively eliminate the false positive signals and the sensitivity limit is0.1%, would be applied to inspection, quarantine and food safety.

关 键 词：酶连接探针杂交芯片 转基因检测 品系鉴定 

分 类 号：TS207.3[轻工技术与工程—食品科学]