摘要
DNA环介导等温扩增(Loop Mediated Isothermal Amplification,LAMP)方法是一种新型的核酸扩增检测方法,该方法操作简便、所需时间短、灵敏度高、特异性强。实时浊度法可以实时检测反应过程中所产生的白色沉淀,从而实现对LAMP整个反应过程的实时监控。本研究以抗草甘膦转基因大豆为研究对象,针对外源基因cp4-epsps的保守区域设计特异性引物,通过实时浊度法在63℃恒温条件下完成转基因大豆的检测工作。结果显示,LAMP实时浊度法能够特异性检测cp4-epsps基因,其检测灵敏度是常规定性PCR方法的10倍。本研究建立了针对转基因大豆cp4-epsps基因的LAMP实时浊度检测方法,该方法具有高度的稳定性与特异性,结果准确,适合于转基因抗草甘膦大豆的快速检测。
Loop-mediated isothermal amplification method （LAMP） is a novel nucleic acid ampli fication technology. The LAMP method amplifies DNA with rapidity, high specificity and sensitivity under isothermal conditions. Since turbidity of the reaction mixture would increase in correlation with the DNA yield, real-time monitoring of the LAMP reaction was achieved by real-time turbidimeter. Enolpyruvl Shimimate phosphate syntheses gene was amplified by a set of four specially primers that recognize six distinct sequences of the target. The amplification can be obtained in 1 h by incubating all of the reagents in a single tube by real-time turbidimeter at 63℃. Results from this study showed that the LAMP method was an effective method for the rapid detection of Transgenic Soybean and their test results were consistent with the results of conventional PCR methods. LAMP assay results were found to be 10 times more sensitive than the conventional PCR. The LAMP detection method was specific, stable and reliable, and will be an effective tool for rapid detection of Transgenic Soybear~
出处
《现代食品科技》
EI
CAS
2011年第10期1264-1267,共4页
Modern Food Science and Technology
基金
广东省质监局科技计划项目(2009zz11)
