As a typical estrogen, 17β-estradiol can easily affect human health through accumulation in animal food. Therefore, the establishment of a simple and sensitive technique for rapid analysis of 17β-estradiol is of great practical significance. On the basis of the gold nanoparticles colorimetric analysis, a gold nanoparticles colorimetric detection method for 17β-estradiol based on aptamer was designed, using the 17β-estradiol aptamer as the sensing probe. The test conditions were also optimized, and the feasibility of this method for the detection of 17β-estradiol in milk and eggs was analyzed. Under the optimized conditions, the A650/A530 of water system showed a good linear correlation with the concentration of 17β-estradiol (R2=0.915) with a linear range from 0.25nmol/L to 0.60nmol/L, and the detection limit was 0.233nmol/L. The proposed method also illustrated an excellent specificity to 17β-estradiol, when the ratio of gold nanoparticles to aptamer was 1∶11000 which were incubated at room temperature for 30min, and under the condition of 24mmol/L NaCl. The method can also be applied successfully for the detection of 17β-estradiol in milk and egg samples. The linear range were 0.5～0.8nmol/L (milk) and 0.3～0.9nmol/L(eggs) with the detection limit of 0.183nmol/L and 0.185nmol/L, respectively. The recovery were 101.6%～105.4% (milk) and 97.97%～115.30% (eggs), respectively. Meanwhile, the relative standard deviation was less than 6%. With the advantage of higher sensitivity and specificity, the simple and practical method can provide a way for the rapid detection of 17β-estradiol in milk and egg samples.