应用MPCR-DHPLC技术检测转基因大豆及其食品

Detection of Transgenic Soybean and Its Food by MPCR-DHPLC Method

利用多重PCR结合DHPLC技术,建立了高通量快速筛选检测转基因大豆及其食品的方法,确立了转基因大豆品系鉴定方法。该筛选方法的检测灵敏度为0.078 ng.μL-1,质粒检测灵敏度为1×103拷贝.μL-1。利用该方法对转基因大豆GT-40-3-2、Mon89788、A5704-12和大豆食品曲奇饼干、含大豆的调味粉、干豆腐及非转基因黑大豆进行验证,效果良好。所建立的PCR-DHPLC检测方法能同时快速准确的检测大豆及加工食品中转基因成分。

The aim of this work was to develop a method for simultaneous detection of a variety of genetically modified（GM）soybean ingredients in foods and to identify the lines of transgenic soybean by using multiplex polymerase chain reaction（PCR）coupled with denaturing high performance liquid chromatography（DHPLC）assay.Results showed that the multiplex PCR-DHPLC method was highly specific for the target genes with 0.078 ng·μL-1 of high sensitivity.Detection sensitivity of standard plasmid was 1×103 copy·μL-1.Identification and detection of genetically modified soybean ingredients in GT-40-3-2,Mon89788,A5704-12,cookies,seasoning powder containing soybean,dried tofu and non-genetically modified black soybean were effective.Multiplex PCR-DHPLC method developed in this work will be a better alternative for the rapid and high-throughput detection of many genetic modification events in soybean and its processed food simultaneously.