This paper presents a sensitive and convenient visual methodology for Salmonella typhimurium detection using gold nanoparticles (AuNPs) as colorimetric probes and magnetic nanoparticles (MNPs) as concentration elements. In the protocol, the aptamers were first immobilized onto the surface of AuNPs and MNPs, respectively. Then, S. typhimurium were added into the above solution and incubated for 45 min. During the incubation, aptamer on the surface of nanoparticles could specifically bind to the target and form a MNPs-aptamer-S. typhimurium-aptamer-AuNPs sandwich structure complex. In a magnetic field, the formed complexes were easily separated from the solution, resulting in a fading of the AuNPs suspension and a decrease of the ultraviolet visible (UV/Vis) signal. The assay shows a linear response toward S. typhimurium concentration through a range of 25 to 10(5) cfu/mL, and the detection limit was improved to 10 cfu/mL. The applicability of the bioassay in real food samples was also investigated; the results were consistent with the experimental results obtained from plate-counting methods. It is believed that the developed aptasensor will broaden the application in bioassays.