The present invention relates to the biologic technology field, and discloses a liquid phase albumen chip and the preparation and the usage method thereof which can simultaneously test human serum carcinoma embryonic antigen (CEA), Alpha fetoprotein (AFP), and hepatitis B surface antigen (HBsAg). The present invention couples the specificity antibody of CEA, AFP, and HBsAg on different fluorescence micro-spheres, and uses the test antibody marked by biotin or phycoerythrin to determine the nature quickly and quantitatively analyze the above three indexes with the double antibody sandwich method. The present invention uses the filtering membrane board when testing, and washes the board 3 times after each reaction is finished to increase the signal and improve the sensitivity. The present invention has high sensitivity, strong specificity, stable result, excellent repeatability, and simple operation; 1 micro liter serum sample can test three indexed simultaneously. The present invention is applicable to the health test and the general examination as well as the clinic test of the high risk population, and can facilitate the early diagnosis and the early treatment of the knub.