Accurate Thrombin Monitoring Based on Proximity Ligation Assay-Assisted Rolling Circle Amplification (RCA)

Due to the fact that the expression level of thrombin affects the coagulation function of the injured tissue after trauma, it is considered as a very promising biomarker for the diagnosis and treatment of trauma. Nonetheless, sensitive, simple, and accurate thrombin detection continue to be extremely difficult. Here, using the two domains of thrombin as detection targets, we build a unique, accurate, isothermal thrombin analysis method. The method is constructed based on the integration of proximity ligation and rolling circle amplification (RCA). This approach specifically binds with the two functional domains of thrombin by using two intricately constructed probes. The technique has great accuracy thanks to proximity ligation, and the coupled RCA ensures acceptable sensitivity. With a limit of detection (LOD) of 0.23 pM, the method has demonstrated favorable detection persistence. Furthermore, the technique has a high selectivity for thrombin. Integrating merits including high sensitivity, low cost, and good portability, this method may enrich the arsenal for thrombin related applications.