Protein-induced fluorescence enhancement as aptamer sensing mechanism for thrombin detection

•A rapid and simple ‘one-pot’ homogenous fluorescence enhancement assay is used to detect thrombin protein.•Applicability of PIFE for thrombin sensing is demonstrated without loss of sensitivity with respect to FRET.•PIFE based thrombin detection displayed excellent linearity in the range of 0.25 pM–25 nM with a detection limit of 8.9 pM.•Developed aptasensor displays high specificity and function in the presence of ‘natural’ backgrounds and various salt conditions.•The proposed approach can be employed as a screening platform for DNA aptamers based protein sensing.