A hybridoma cell line of secreting clarithromycin monoclonal antibodies with a preservation number of hybridoma cell line of CGMCC No. 14696 belongs to the field of food safety immunological detection. BALB/c mice are immunized through one time immunization with complete freund's adjuvant, three times of booster immunization with incomplete freund's adjuvant and one time of rush immunization with clarithromycin complete antigen without adjuvant; the spleen cells from BALB/C mice immunized with high potency and low value of IC50 are fused with murine myeloma cells; and then the hybridoma cell line is obtained through indirect competitive ELISA screening and three subclones. The monoclonal antibody secreted by this cell line has good specificity and detection sensitivity to clarithromycin (value of IC50 is 0.3 ng/ml), being suitable for detection of clarithromycin in food.