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Specific primers and blockers were designed based on Cronobacter sakazakii gene (16SrRNA) and Real-time Fluorescence Single Primer Isothermal Amplification (RF-SPIA) was established to detect C. sakazakii. This method identified that C. sakazakii genes were positive, whereas other non-C. sakazakii genes were negative. The sensitivity of this test for C. sakazakii pure culture reached 1.4×100CFU/mL and that for C. sakazakii genome reached 7.5×101fg/mL. The detection limit of RF-SPIA in milk powder artificially contaminated with C. sakazakii could reach 8.1×101CFU/mL. These results demonstrate that the RF-SPIA detection method for C. Sakazakii is highly sensitive, strong, specific and convenient.

Detection of <em>Cronobacter sakazakii</em> in Powdered Infant Formula using Real-time Fluorescence Single Primer Isothermal Amplification