普通PCR法、荧光定量PCR法及微流控芯片法检测大豆产品中外源基因灵敏度的比较  

Comparison on Sensitivitys of PCR, Real-time PCR and Microfluidic Chip for Detection of Foreign Gene in Soybean Products

作　　者：吴姗[1,2] 吴志毅 张晓峰 黎昊雁 

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》2009年第2期176-186,共11页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家质检总局科技计划项目(ZK200512)

摘　　要：目的:为提高食品中转基因成分检测的灵敏度,将微流控芯片仪应用到转基因检测中。方法:将微流控芯片法与普通PCR法、荧光定量PCR法进行灵敏度比较;检测高温、高压处理过的转基因大豆粉。结果:荧光定量PCR法的灵敏度明显高于普通PCR法,但在较极端高温、高压处理(高压灭菌121℃/30 min)及转基因成分含量较低(1%)的情况下,采用荧光定量PCR法未能检测到外源基因,而微流控芯片法能检测到相应的峰值,前提是在未添加内标(Marker)的情况下。若添加内标,在内标峰和目的峰区分开的条件下,有损检测灵敏度。结论:微流控芯片在转基因检测中具有高效率、高灵敏度的特点,但该方法有待完善。Objective： To improve the sensitivity of detection for transgenic component in foods, mierofluidic chip was emploied into detection of foreign gene in soybean. Methods： The microfluidic chip, PCR and Real-time PCR also have been applied to detect foreign genes in the complicated material, the transgenic soybean which was ground into flour and subjected to autoclave, to simulate the condition of high temperature and high pressure in the course of food process. Results： By comparison, in sense of sensitivity, Real-time PCR was significantly higher than PCR. However, more extreme treatment, when autoclaved at 121 ℃/30 min, and GMO content lower to 1%, Real-time PCR failed to detect foreign genes. At the same time, microfluidic chip was able to detect the corresponding peak, but in the circumstances of no internal standard （Marker）. If adding the internal standard, it is necessary to distinguish between internal standard and purpose summit; and under that condition, generally, the detection sensitivity of microfluidic chip would be undermined. Conclusion： Microfluidic chip is very efficient and sensitive in detection foreign gene in GMO. but this method have also many problems which remain to be solved.

关 键 词：转基因检测 普通PCR 荧光定量PCR 微流控芯片 

分 类 号：S565.1[农业科学—作物学]