基于微滴式数字PCR技术，建立了转基因玉米MON87411双重微滴式数字PCR定量检测方法。试验结果显示，该方法仅有玉米MON87411能特异性检出，其他样品均无扩增反应。扩增稳定性试验，3次平行扩增内外源基因拷贝数RSD分别为0.56%和3.93%，均符合RSD小于25%的要求。线性相关性曲线显示，当模板DNA质量浓度在50~0.08 ng/μL之间时，目标序列拷贝数与模板DNA浓度具有良好的线性关系，相关系数R2均大于0.99，品系特异性序列定量线性范围在26.6~17 086.6拷贝之间，定量限（LOQ）为1.34拷贝/μL。准确度试验，五个不同质量浓度模板DNA平均拷贝数含量分别为52.14%、63.09%、66.08%、60.50%、52.16%，均在理论拷贝数含量范围内。综上所述，该研究建立的双重微滴式数字PCR定量检测方法特异性强，稳定性好，定量范围广，灵敏度和准确度高，可以用于转基因玉米MON87411的定量检测。

Based on the droplet digital PCR technology, a quantitative duplex droplet digital PCR detection method for genetically modified maize MON87411 was established. The results showed that only the maize MON87411 could be specifically detected by this method, with no amplification reaction being observed in other samples. In the amplification stability test, the copy number RSD of endogenous and exogenous genes in three parallel amplifications were 0.56% and 3.93%, respectively, which met the requirement of RSD less than 25%.The linear correlation curve showed that when the mass concentration of template DNA was between 50 ng/μL and 0.08 ng/μL, there was a good linear relationship between the copy number of target genes and the template DNA concentration, with correlation coefficients R2 greater than 0.99. The quantitative linear range of MON87411-specific gene was between 26.6 and 17 086.6 copies with the limit of quantification (LOQ) being 1.34 copies/μL. In the accuracy test, the average copy number content at the five different template DNA mass concentrations were 52.14%, 63.09%, 66.08%, 60.50% and 52.16%, respectively, which were all within the theoretical copy number content range. In summary, the quantitative duplex droplet digital PCR detection method established in this study had high specificity, good stability, wide quantification range, high sensitivity and accuracy, thus can be used for the quantitative detection of genetically modified maize MON87411.

科技部科技基础资源调查专项（2023FY100402）；海关总署科研项目（2024HK044）；拱北海关科研项目（2021GK010）