Sensitive detection of African swine fever virus p54 based on in-situ amplification of disposable electrochemical sensor chip

Abstract

Sensitive detection of African swine fever virus (ASFV) is important to reduce the loss of pig industry. A highly sensitive disposable electrochemical sensor chip for the detection of ASFV p54 was developed by combining in-situ loop-mediated isothermal amplification (LAMP) with electrochemical sensing technology. The disposable electrochemical sensor chip was prepared by screen printed carbon electrode (SPCE) covered with polydimethylsiloxane membrane as micro-electrolytic cell, and the primers designed according to the virus were fixed on the gold nanoparticles (Au NPs) modified SPCE. The target gene was amplified by in-situ LAMP within 42 min, and the deposition of Au NPs can promote the electron transfer, enhance the electrochemical signal of [Fe(CN)6]3-/4- probe and improve the sensitivity. The detection range of the biosensor was 1.0 × 10-12-1.0 × 10-6 g/L, the detection limit was 3.0 × 10-13 g/L (0.48 copies/µL). In addition, the sensor has good results for the detection of real samples and has broad application prospects.