大豆深加工产品中外源基因检测灵敏度的研究  

Studies on the Sensitivity of Detection for Foreign Gene in Deep-processed Soybean Products

作　　者：吴姗[1] 吴蓉[2] 林晓佳 吴志毅 

机构地区：[1]浙江出入境检验检疫局,杭州310012 [2]中科院上海植物生理生态研究所,上海200032

出　　处：《中国食品学报》2008年第4期139-146,共8页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家质检总局科技计划项目(No.ZK200512)

摘　　要：为了提高大豆深加工食品中外源基因的检测灵敏度,改良了普通PCR反应的引物,并设计了NEST-PCR反应程序。模拟食品加工中高温、高压等处理方法,将转基因大豆研磨成粉状,在高压灭菌锅内进行高温、高压处理。应用新设计的引物35SCP142、CPNOS165进行外源基因检测,通过对引物的调整,检测灵敏度得到了明显的提高,可以检测到的转基因大豆含量仅为1%,且为经过121℃/30min处理的样品中的外源基因。应用NEST-PCR的高灵敏度的特性,设计了NEST-PCR的两对引物35SCP318和35SCP114,并检测到外源基因的片段。将普通PCR的检测灵敏度和荧光定量PCR的灵敏度作比较,若引物设计合理,且PCR条件适合,普通PCR的检测灵敏度可以和荧光定量PCR的相媲美。In order to improve the sensitivity of detection for foreign genes in deep-processed soybean products, primers applied in common PCR were optimized and the protocol of NEST-PCR were designed. The transgenic soybean was ground into flour and subjected to autoclave, to simulate the condition of high temperature and high pressure in the course of food process. With the new designed primers 35SCP142 and CPNOS165, sensitivity of detection the foreign genes in deep-processed （or autoclaved） soybean flour has been obviously improved by optimizing the primers. In the sample treated with autoclave （121 ℃/30min） and transgenic soybean content only 1%, the foreign genes also could be detected with the new primes. Another two primers 35SCP318 and 35SCPl14 for NEST-PCR have also been designed, with which DNA fragments of foreign gene has been amplified and detected. Compared two methods common PCR and fluorescent quantitation PCR, if design of primer is rational and PCR conditions are also suitable, the detection sensitivity of common PCR and fluorescent quantitation PCR could be comparable.

关 键 词：转基因检测 引物优化 NEST—PCR 荧光定量PCR 

分 类 号：TS214.2[轻工技术与工程—粮食、油脂及植物蛋白工程]