Cas-mCfLAMP: A multiplex rapid visualization assay for sugarcane pathogens based on labeled LAMP and CRISPR/Cas12a

Sugarcane diseases are caused by various pathogens, and rapid and accurate identification of these diseases has many challenges. Detection of sugarcane diseases using conventional assays are often handicapped by their lowthroughput, high-cost, and low-sensitivity nature. Here, we report a novel method for the multiplexed and visual detection of sugarcane pathogens using CRISPR/Cas12a coupled with labeled loop-mediated isothermal amplification (Cas-mCfLAMP). This method labels the product during LAMP amplification, relieving the high dependence of CRISPR/Cas12a on specific CrRNAs and enabling the detection of multiple target genes in a single reaction. In addition, we have developed a field detection system that allows visual detection of sugarcane diseases in the field without the need for specialized technicians or expensive equipment. This method is simple, efficient, specific, and cost-effective for sugarcane disease detection, and holds potential to be modified for the field detection of other plant diseases.