Field-Deployable Detection of Genetically Modified Organisms with an Integrated Method of Loop-Mediated Isothermal Amplification and CRISPR/FnCas12a.

The detection of genetically modified organisms (GMOs) is crucial for regulatory compliance and consumer safety. This study presents a novel method combining loop-mediated isothermal amplification (LAMP) with CRISPR/Cas12a cleavage, termed Cas-pfLAMP, for sensitive and specific GMO detection. We developed assays for three GM events: maize DBN9936 and MON810 and soybean GTS40-3-2. By incorporating a universal protospacer adjacent motif (PAM) sequence into LAMP primers, we overcame the limitations of PAM site dependence. The Cas-pfLAMP assays demonstrated high specificity and sensitivity, with limits of detection as low as 10-12 copies per reaction. Furthermore, we developed a point-of-care testing platform integrating rapid DNA extraction, Cas-pfLAMP, and lateral flow strips for on-site GMO detection. This platform achieved comparable sensitivity to qPCR, detecting GM contents as low as 0.1% in simulated samples within 40 min. The Cas-pfLAMP method offers the advantages of PAM site independence, high specificity and sensitivity, and suitability for field testing without specialized equipment. This approach represents a promising new generation of GMO detection methods with potential applications in various scenarios.