As a food adapted to all kinds of people, milk has a high nutritional value. Because milk is a complex biological matrix, detecting illegal compounds is often difficult. As a common pesticide, organophosphorus (OP) residues caused by nonstandard use may be ignored, which is a threat to milk quality. In this study, using coumaphos as template molecule, the synthesized molecularly imprinted polymer (MIP) can specifically recognize 7 kinds of OP. Then, the MIP was used as an identification element to prepare a chemiluminescence sensor on a 96-well microplate for the determination of OP residues in milk samples. Due to the 4-(imidazol-1-yl)phenol-enhanced luminol-H 2 O 2 system, the sensitivity of the system is very high; the detection limits of 7 OP including coumaphos, fenthion, chlorpyrifos, parathion, diazinon, fenchlorphos, and fenitrothion were 1 to 3 pg/mL, and the half maximal inhibitory concentrations were 1 to 20 ng/mL. The intraday recoveries of 7 OP were in the range of 86.1 to 86.5%, and the interday recoveries were in the range of 83.6 to 94.2%. Furthermore, the sensor can be reused up to 5 times. Therefore, the MIP-based chemiluminescence sensor can be used as a routine tool to detect OP residues in milk samples.