摘要
根据外源插入序列与大豆内源基因边界序列,用设计软件Primer Explorer Version 3设计并筛选了转基因大豆GTS 40-3-2品系的环介导等温扩增引物,并进行了反应体系和反应条件的优化,建立了转基因大豆GTS 40-3-2品系特异性环介导等温扩增检测方法。对该方法进行了特异性、灵敏度、稳定性评价,结果显示:该方法能够特异、稳定、灵敏地检测大豆及其制品中的转基因大豆GTS 40-3-2成分,检测低限达到0.5%。此外,对12份实际样品的检测结果表明,该方法与实时荧光PCR方法的检测性能相当,结果吻合率为100%,假阳性率和假阴性率均为0。
According to the exogenous insert sequences connected with the border sequences of soybean endogenous genes, we designed and screened specific loop-mediated isothermal amplification primers, and optimized the reaction system and the reac- tion conditions using a design software named Primer Explorer Version 3,then established the specific loop-mediated isothermal amplification method for the detection of the transgenic soybean line GTS 40 - 3 - 2 strains. The specificity, stability and sensi- tivity of this method were evaluated. The results indicated that the transgenic soybean GTS 40 - 3 - 2 could be distinguished from various soybean strains and their derived products specifically,stably and sensitively with LOD of 0.5 %. In addition, the detection results of 12 samples were 100% consistent to that of the real-time PCR method, with the zero false positive and nega- tive rates,which indicated that this method was equivalent to the real-time PCR method.
出处
《粮食与饲料工业》
CAS
北大核心
2012年第12期60-63,共4页
Cereal & Feed Industry
基金
出入境检验检疫行业标准制定计划项目:出口食品中转基因成分环介导等温扩增(LAMP)检测方法(2011B286k)
广东省科技计划项目(2009B0627):食品转基因成分快速检验能力建设(粤科函财字[2009]627号)
广东检验检疫局科技计划项目:主要转基因食品的快速筛查技术及装置研究(2009GDK45)
