Objective: The development of a method for detecting Cronobacter spp in milk powder based on CRISPR/Cas12b. Methods: Firstly， the amplification primers and guide RNAs were designed and synthesized based on the specific gene sequence of Cronobacter spp. The conditions， specificity and reaction system were optimized， and the sensitivity and reproducibility of the system were validated using artificially contaminated samples. Results: The results showed a high specificity and sensitivity of this method - the sensitivity reached 0.01 ng/μL； samples with initial bacterial counts less than 10 CFU/g could be detected； and the detection limit was<10 CFU/100 g. 51 commercial milk powder samples were later tested using our established method， and the results were consistent with those obtained by traditional national standard method. Conclusion: As such， the CRISPR/Cas12b method established in this study can be used for rapid detection of Cronobacter in milk powder.