绿豆源基因的实时荧光定量LAMP检测方法研究

Study on the Real-time Fluorescence Loop-mediated Isothermal Amplification of Mung Bean-derived Gene

针对绿豆转录间隔区设计一套引物,通过温度优化、特异性和灵敏度试验,建立一种绿豆源DNA的实时荧光定量环介导等温扩增(LAMP)检测方法。结果显示,实时荧光定量LAMP检测方法在温度为61℃时,引物能够将绿豆、大豆、玉米、木薯和马铃薯DNA区分开,灵敏度可以达到0.1pg·uL-1。实时荧光定量LAMP方法能够快速有效地对绿豆源成分进行检测,为绿豆制品真实性检测提供了一种快速准确的检测方法。

To establish a real-time fluorescence loop-mediated isothermal amplification(lamp) method for the detection of mung bean-derived gene, LAMP primer was designed by the internal spacer(ITS) of the specific mung-bean DNA to be used for optimizing the reaction temperature, specificity, and sensitivity. It was founded in the study that the optimal temperature of the LAMP method was 61℃. Under the reaction conditions of 61℃, the detection limit of LAMP was 0.1 pg uL-1 with the good primer specificity for the mung bean DNA which could be distinguished from the DNAs of soy bean, corn, cassava, and potato. LAMP which has the advantages of simple operation, strong specificity and high sensitivity could effectively detect the mung bean-derived gene. Therefore, the real-time fluorescence lamp method could be used as a rapid detection method for the adulteration of mung bean products.