Conventional magnetic relaxation switching (MRS) sensor suffers from its relatively low sensitivity when it comes to the analysis of trace small molecules in complicated samples. To meet this challenge, we develop a cascade reaction-mediated magnetic relaxation switching (CR-MRS) sensor, based on the assembly of silver nanoparticles (Ag NPs) and magnetic nanoparticles (MNPs) to improve the sensitivity of conventional MRS. The cascade reaction triggered by alkaline phosphatase generates ascorbic acid, which reduces Ag+ to Ag NPs that can assemble the initially dispersed MNPs to form magnetic/silver nanoassemblies, thus modulating the state of MNPs to result in the change of transverse relaxation time. The formed magnetic/silver nanoassemblies can greatly enhance the state change of MNPs (from dispersed to aggregated) and dramatically improve the sensitivity of traditional MRS sensor, which makes this CR-MRS sensor a promising platform for highly sensitive detection of small molecules in complicated samples.