A novel analysis method based on specific recognition of aptamer and fluorescence probe technique was established for the detection of total residues of tetracycline antibiotics(TCs)，including oxytetracycline,tetracycline,chlortetracycline and doxycycline etc.in aquaculture wastewater.Two segments of DNAs work together to identify TCs to fold into a stable “hairpin” double chain structure that can be inserted by nucleic acid dye DAPI to generate of fluorescence emission.Based on this,TCs can be quantified.Under the optimal experimental conditions,the working curve was established with oxytetracycline as target molecule,the fluorescence intensity showed a good linear relationship with concentration of oxytetracycline in the range of 10-50 nmol/L,and the detection limit of oxytetracycline was 2.3 nmol/L.When having oxytetracycline,tetracycline,chlortetracycline and doxycycline as additives,the average recoveries ranged from 88.5% to 102.3%,with relative standard deviations(n=5) of 3.2%-6.7%.The method is simple,sensitive and rapid,and could satisfy the requirements for determination of total TCs residues in the farm wastewater.