A Highly Sensitive Electrochemiluminescence Biosensor for Pyrophosphatase Detection Based on Click Chemistry-Triggered Hybridization Chain Reaction in Homogeneous Solution

The abnormal expression of pyrophosphatase (PPase) is closely related to many diseases and malignant tumors, so the detection for PPase is of great significance in clinical diagnosis, disease monitoring, and other biomedical aspects. In this study, a sensitive and specific electrochemiluminescence (ECL) biosensor combined highly specific Cu+-catalyzed azide-alkyne cycloaddition (CuAAC) with high efficiency of hybridization chain reaction (HCR) for the purpose of detecting pyrophosphatase has been designed. Highly efficient hybridization chain reaction amplification processed in homogeneous solution and the amplification products were connected to the electrode surface in one step, which solved the problem of low DNA amplification efficiency on the electrode surface because of the steric hindrance. Ru(phen)32+ was embedded into the dsDNA and functioned as ECL probes; the enhanced ECL intensity of the system had a linear relationship with the logarithm of PPase concentration in the range of 0.025–50 mU with a detection limit of 8 μU. The method was proved to be of good specificity, repeatability, and stability that could be used for screening and quantitatively determining pyrophosphatase inhibitor sodium fluoride. The practicability of this method in clinical application has been proved through the detection of serum from the clinical arthritis patients. Moreover, the method can be used to monitor PPase activity of arthritis patients before and after administration to provide reference for the effect of drug treatment.