In this paper, a novel apta-sensing colorimetric platform for rapid detection of chloramphenicol (CAP) in raw milk was developed. The AuNPs are stabilized by short-sequences aptamers against salt induced aggregation and this is the base of most colorimetric aptasensors development. However, the statute shows low sensitivity for the long-sequence aptamers. Herein, we propose an alternative strategy that use intact long-sequence aptamers for develop a highly sensitive AuNP-based colorimetric aptasensor. Determination of CAP in animal derived foods is an urgent demanded in the effort to minimize food safety risk. Therefore, we chose it as the representative model to construct the colorimetric sensing platform based on aptamer terminal-lock (ATL). In the ATL, intact aptamer was used as a molecular recognition element and a short-sequence oligonucleotide serving as a locker probe (LP) which is complementary of aptamer terminal fragments. By formation of aptamer/target complex, the LP leaves the ATL and adsorbs on the surface of AuNPs, leading to the AuNPs stabilization against salt-induced aggregation. This aptasensor shows a low limit of detection (0.03 nM) with high selectivity toward CAP. Moreover, the designed sensing platform was successfully applied to detect CAP in the milk samples. These results demonstrate our introduced label-free method for CAP detection is simple, sensitive, and highly selective.