转基因植物Real-time PCR方法检出限和定量限的研究

目前Real-timePCR(Rt-PCR)方法在转基因植物定量检测中应用最为广泛。有关该方法的检出限和定量限的计算并没有达成统一。本文采用统计模型对三种转基因植物进行实时荧光定量PCR方法检出限和定量限的研究。实验结果表明:转基因玉米NK603检出限5拷贝,定量限14拷贝;转基因棉花Mon15985检出限5拷贝,定量限12拷贝;转基因油菜Oxy235检出限4拷贝,定量限9拷贝。是利用统计学方法对转基因植物荧光定量PCR方法检出限和定量限研究的一个积极尝试,可用于其他转基因植物检测中检出限和定量限的确定。

Study on limit of detection and limit of quantification of Real-time fluorescence quantitative PCR in gene modified plant

At present,Real-time PCR(Rt-PCR)is the most widely used in the quantitative detection of gene modified plant.Calculation of the limit of detection(LOD)and limit of quantification(LOQ)in this method was not obtained consistency.The LOD and LOQ of real-time fluorescence quantitative PCR in three kinds of gene modified plants were studied by statistical model.The results showed that LOD and LOQ of transgenic maize NK603 were 5 copies and 14 copies,respectively.LOD and LOQ of transgenic cotton Mon15985 were 5 copies and 12 copies,respectively.LOD and LOQ of transgenic canola Oxy235 were 4 copies and 9 copies,respectively.An active try on LOD and LOQ of transgenic plant was studied by statistical method,which could be used to determine the LOD and LOQ of other gene modified plants.