"Blue-red-purple" multicolored lateral flow immunoassay for simultaneous detection of GM crops utilizing RPA and CRISPR/Cas12a.

Advanced multiplexed testing techniques should be designed and developed to ensure an accurate and reliable evaluation for unknown samples. In this study, an efficient platform coupled with the "Blue-Red-Purple" strategy based on recombinant polymerase amplification (RPA), CRISPR/Cas12a and lateral flow strip was established, which could realize the dual-target detection of CP4-EPSPS and Cry1Ab/Ac in genetically modified crops. The lateral flow immunoassay was developed using different colored microspheres to label the antibodies to realize the visualization of results and avoid cross-reactions. The proposed method exhibits high specificity, sensitivity and stability. The visual detection limits of standard plasmids and real samples reached 10 copies/μL and 0.5 %, respectively, which could be stored at 4 °C for 12 months with high detection ability. Moreover, the entire detection process could be completed within 50 min without any complex instruments or professional operators. These findings indicated that a sensitive, specific, rapid and accurate method was established for on-site detection of GM crops.