The invention relates to the technical field of
food safety molecule detection, in particular to a method and a device for detecting food-borne pathogenic microorganisms by using a tubular device based on a Cas12a
system, the tubular device is divided into an upper layer and a lower layer according to a mechanical structure, an RPA reaction
solution system enrichment target is placed on the upper layer, a
CRISPR-Cas12a
system is placed on the lower layer, so that physical isolation is formed, and the Cas12a
system is arranged on the lower layer. The upper-layer reaction is independently completed. The RPA product enters the lower layer through a gap by moving the tube cover, Cas12a and crRNA recognize and are specifically combined with a target, so that Cas12a trans-cleavage activity is activated, ssDNA marking a
fluorophore HEX and a
quenching group BHQ1 in a system is cleaved, a
fluorescence signal is released, and a result can be observed through a
blue light lamp and a filter. Sample detection can be completed within 40 min, the cost is low, operation is easy and convenient, system
assembly can be completed at a time in the detection process, repeated uncovering is not needed, the requirement for equipment is low, and the method can be suitable for on-site
rapid detection.