The invention relates to a magnetic particle chemiluminescence double-layer micro-fluidic chip used for whole-blood sample detection. The micro-fluidic chip comprises a top plate (1) and a bottom plate (2). The top plate comprises a sample feeding port (4), a sample filling region (12), a marked ligand storage pool (5) and a sample mixing region (13). The bottom plate comprises a filtering region (6), a magnetic particle coating region (7), a cleaning region (14), a detecting region (8), a cleaning fluid storage pool (9), a luminous substrate liquid storage pool (10) and a liquid release channel (16).