The invention belongs to the technical field of environmental
biological safety monitoring, and particularly relates to an RPA-
CRISPR method for rapidly detecting pathogens in air. According to the invention, a two-
step detection strategy integrating RPA isothermal amplification and
CRISPR detection is established; the method specifically comprises the following steps: firstly, acquiring
genome DNA of an
air sample, and carrying out specific amplification on a trace target by utilizing an RPA technology; then, a product is introduced into a
CRISPR-Cas12a
system, and the trans-shear activity is recognized and activated by utilizing the specificity of crRNA, so that
signal cascade amplification is realized.
Pseudomonas aeruginosa is used as a model
pathogen, and a primer and probe combination is designed aiming at a specific
gene of the
pseudomonas aeruginosa. Results show that the method can complete detection within 10-15 minutes at the soonest, the lowest
detection limit can reach a
single copy level (100 copies / L), and the method has the advantages of high environmental
interference resistance, high sensitivity and no need of large instruments, and can meet the on-site rapid screening requirements of air
pathogen aerosol in public places, medical environments and other scenes.