超分枝滚环扩增技术结合试纸法检测食品中多种转基因组分

HRCA-based Strip Test for Detection of Transgenic Components in Food

目的:建立一种基于超分枝滚环扩增技术的试纸检测方法,用于对食品中存在的转基因成分进行定性检测,以实现多组分同时检测。方法:以转基因和非转基因大豆、转基因玉米、转基因马铃薯按比例混合配制的多组分转基因复合材料为研究对象,考察固定有特异性检测探针的硝酸纤维素膜试纸,能否有效检出经连接和滚环扩增获得的反应产物中的转基因组分。结果:3种转基因组分中,无论是采取单组分,还是两两混合或3种同时混合,固定有特异性检测探针的硝酸纤维素膜试纸均可将其检出,准确性为100%;试纸可从基因组DNA中检出100pg/μL的转基因组分,灵敏度满足常规检测要求。结论:初步建立了多组分转基因成分试纸检测方法,其准确性和特异性能够满足常规检测要求。

This study aimed to develop a strip test method based on HRCA technique for simultaneous qualitative detection of various transgenic components in food.A mixture of transgenic and nontransgenic soy bean and transgenic maize and potato in some proportion was prepared to evaluate the effectiveness of nitrocellulose membrane strips developed to immobilize specific probes in detecting transgenic components in reaction products obtained through linkage and rolling-cycle amplification.Results showed that nitrocellulose membrane strips carrying specific probes achieved 100% accuracy in detection of pairwise mixtures or full mixture of the above transgenic components or each of them and gave a detection level of 100 pg/μL,which meets the requirements of routine detection.The developed nitrocellulose membrane strips carrying specific probes may provide an effective tool for detection of transgenic components in food.