Sensitive detection of alkaline phosphatase by dephosphorylation-initiated transcription reaction-mediated dual signal amplification

Abstract

We develop a new fluorescence method for the sensitive detection of alkaline phosphatase (ALP) based on dephosphorylation-initiated transcription reaction-mediated dual signal amplification. This research demonstrates for the first time a significant improvement in ALP assay performance by using a nucleic acid amplification strategy, which enables sensitive detection of ALP activity in vitro and in cell extracts. Importantly, this method can be further applied for kinetic analysis and ALP inhibition assay.