Isothermal protein detection assay based on high efficient T7 RNA polymerase transcription-mediated amplification system and graphene oxide fluorescence switch platform

An isothermal protein detection assay based on high efficient T7 RNA polymerase transcription-mediated amplification system and graphene oxide (GO) fluorescence switch platform is developed. The assay consists of specific aptamer-protein recognition, high efficient T7 RNA polymerase transcription-mediated amplification of DNA aptamer, convenient and rapid GO fluorescence switch platform for the detection of ssRNA transcripts. Thrombin from human plasma is used as an example to demonstrate the feasibility of the assay. Experimental results show that the detection limit of the assay is 10 pM of thrombin. A calibration curve with a linearity range from 10 pM to 10 nM is established, thus, make quantitative analysis possible. The assay can specific discriminate thrombin from other proteins. It holds great promise in protein detection due to its sensitivity and specificity.