The invention discloses a micro-fluidic chip immunoassay kit and a detection method thereof. The kit comprises a micro-fluidic chip, a wafer cover plate and a wafer bottom plate, wherein the wafer cover plate and the wafer bottom plate are bonded up and down. One or more detection units are arranged in the wafer bottom plate, and each detection unit comprises a liquid storage tank, a mixed reaction tank, a detection tank, a waste liquid tank and a first vent hole which are connected in sequence; the mixed reaction tank is fixedly provided with a fluorescent probe corresponding to a detection drug; goat anti-mouse immune globulin G and corresponding drug antigens are fixed in the detection tank. The invention also discloses a detection method using the kit. The method has the characteristics of high flux, low sample consumption, high detection speed, simplicity and convenience in operation and the like; the method has the advantages of no need of an electrode, a voltage or an external magnetic field, no need of electroosmosis driving, hot air pump driving or optical capture micropump driving, simultaneous detection of various veterinary drug residues in various matrixes, suitableness for on-site rapid screening analysis, and great alleviation of the detection pressure of related detection personnel.