Establishment of real time fluorescence single primer isothermal amplification method for detection of Cronobacter sakazakii.Methods Taking Cronobacter sakazakii OmpA gene specific sequence as the target sequence, the RNA-DNA combination primer and chain termination sequence was designed, and the reaction system was optimized. 4 different strains of Cronobacter sakazakii strains and 21 other foodborne pathogens were detected by the method, and the specificity of the real-time fluorescent SPIA was analyzed.The sensitivity was analyzed by different dilution of Cronobacter sakazakii and the detection limit was determined by spiked infant formula. Results Only Cronobacter sakazakii could be detected and showed the typical fluorescence curve. The method has good specificity. In 40 min, the sensitivity of real-time fluorescent SPIA for the detection of Cronobacter sakazakii in pure culture was 1 copies/reaction. The corresponding number of the living bacteria was 1.6×10-1 cfu/ml. The detection limit of Cronobacter sakazakii in infant formula is 1.5×10⁰ cfu/100 g.Conclusion The method has the advantages of high sensitivity, strong specificity, less time-consuming.