The long-lived radioisotope 59Ni is of interest in various research fields including neutron dosimetry, radioactive waste management, and astrophysics. In order to achieve the sensitivity required for such applications, the technique of accelerator mass spectrometry (AMS) 59Ni measurement has been developed at the AMS facility at China Institute of Atomic Energy (CIAE). Based on the AE-Q3D detection system in the CIAE AMS facility, the interference in 59Ni counting from the isobar 59Co has been reduced by a factor of 8 × 106. A series of laboratory reference samples and a blank sample were measured to check the performance of 59Ni measurement. A detection sensitivity of about 5 × 10−13 (59Ni/Ni) has been obtained.