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Highlights • Structure-switching aptamer probe was designed to achieve enzyme-free amplification. • Detection process for AFB 1 could be finished within 1 min. • All detection process could be conducted in one test-tube at a constant temperature. • AFB 1 contamination in fluid or liquid food samples were accurately detected. Abstract Aptamer probes provide an opportunity for achieving rapid and on-site detection of food contaminants. Herein, we proposed a general design strategy for aptamer probes enabling enzyme-free amplified, ultrafast and one-test tube homogeneous detection of aflatoxin B 1 (AFB 1). The key feature of the aptamer probe is designed with dual-terminal proximity structures, allowing the binding of one molecule to light up two fluorophores, leading to enzyme-free amplification and a remarkable improvement of signal to background ratio and sensitivity for AFB 1 detection. This aptamer probe could accommodate quick response to AFB 1 , and the detection process could be finished within 1 min, ranking one of the quickest assays for AFB 1. AFB 1 detection of broad bean paste and peanut oil conferred satisfactory recoveries ranging from 90.3% to 114.8%. Contributed to the generality and simplicity of the design strategy, this structure-switching probe could potentially act as a general platform of on-site detection for food safety. [ABSTRACT FROM AUTHOR]