In this study, a cascade screening system has been developed combining Dual Super Polymerase Chain Reaction (DSPCR) with the universal Lateral Flow Biosensor (LFB) for the ultrafast, universal and visual screening of dual GM elements, taking P-35s × T-nos for example. In the design of DSPCR for universal screening, gene-specific forward primers were labelled with biotin and gene-specific reverse primers were tagged with Cy5 and digoxin, respectively. In 2.5-min, DSPCR effectively amplified the dual target fragments through our prototype facility. Then, through specific antigen-antibody binding, a universal lateral flow biosensor exported visually dual-amplified results simultaneously without cross contamination. After optimization, the detection limit allowed 0.05% GM maize, corresponding to nine copies in maize. The entire detection process could be achieved in 10 min without any large-scale instrumentation. This method may be useful for the ultrafast, universal and visual screening of dual GM elements (P-35s × T-nos) in GM crop lines and is expected to be of great promise for rapid GMO screening and point-of-care tests.

在这项研究中，级联筛选系统已经发展组合d UAL小号UPER P olymerase Ç海恩ř反应的影响（DSPCR）与通用大号ateral ˚F低乙iosensor（LFB），用于超高速，双GM元件的普遍和视觉筛选，以P- 35s  ×T- nos例如。在用于通用筛选的DSPCR设计中，分别用生物素标记基因特异性正向引物，并分别用Cy5和地高辛标记基因特异性反向引物。在2.5分钟内，DSPCR通过我们的原型设备有效扩增了双重目标片段。然后，通过特异性的抗原-抗体结合，通用的侧向流生物传感器可同时输出视觉上双放大的结果，而不会产生交叉污染。优化后，检出限允许0.05％的转基因玉米，相当于玉米中的9个拷贝。无需任何大型仪器，即可在10分钟内完成整个检测过程。此方法可能对双GM元素（P- 35s  ×T- nos）在转基因作物品系中，有望为快速的GMO筛选和即时检验提供广阔的前景。