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

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.