Dual signal amplification coupling with DNA-templated silver nanoclusters for sensitive and label-free detection of thrombin

Sensitive and reliable determination of thrombin is relevant in the realms of medical and biological research as it serves as an essential biomarker of a number of blood-related illnesses. Herein, we integrate allosteric probe-based specific identification of thrombin and dual signal amplification to present an unique fluorescent technique for label-free and sensitive thrombin detection. Based on DNA polymerase and endonuclease-assisted signal amplification, the method exhibits a high sensitivity with a low limit of detection of 2.3 pM, while maintaining an excellent selectivity and stability. More importantly, the approach is successfully applied in analyzing the effect of nalbuphine on coagulation function of mice. Overall, this approach possesses the advantages of high specificity and sensitivity in label-free detection of thrombin, which is promising in the diagnosis of blood-related diseases.