Abstract: In order to rapidly and conveniently detect Enterobacter sakazakii in milk, a new method was developed by loop-mediated isothermal amplification (LAMP) . The LAMP primers were designed on the basis of outer membrane protein A gene (OmpA) in Enterobacter sakazakii. The LAMP reaction mixture was optimized. The determination limit of LAMP when testing E. sakazakii templates in pure culture was 3.7×101cfu/mL, and its sensitive was better than PCR. For dairy samples, LAMP could specifically detect E.sakazakii cells as low as 4.3×101cfu/mL. Only four Enterobacter sakazakii strains were amplified using LAMP, and no amplicon was observed in other 19 bacteria strains. Therefore LAMP method was an effective technology with high specificity and sensitivity, to conveniently detect E. sakazakii in milk.