A commercial chromogenic agar medium (DFI) was supplemented with glucose (mDFI) to enhance the specificity of Enterobacter sakazakii (E. sakazakii) detection. Escherichia vulneris (E. vulneris), a putative false-positive strain on the DFI medium, produces alpha-glucosidase. The enzyme alpha- glucosidase hydrolyzes a substrate, 5-bromo-4-chloro-3- indolyl-alpha,D-glucopyranoside (XalphaGlc), producing green colonies. E. sakazakii strains produced green colonies on both DFI and mDFI agar, whereas E. vulneris produced green colonies on DFI agar but small white colonies on mDFI agar. E. sakazakii and E. vulneris were also readily differentiated by colony color when the mixed culture of the two strains was plated on mDFI agar and incubated for 24 h at 37 degrees C. The results indicate that the selectivity of the commercial chromogenic agar medium could be improved by a simple supplementation with glucose.