Smart Regeneration Strategy Featured with On-Chip Programmable Thermal Profile for Renewable Aptasensing of Norovirus

Aptamer (Apt)-based biosensors are promising tools for resource-limited Norovirus (NoV), where GII.4 is the predominant sub-genotype causing human infections. Considering their urgent and strict needs of on-site renewal, we present a smart regeneration strategy, inspired by the modern electronics industry, on a homemade screen-printed electrode (SPE). Aqueously prepared MXene@MWCNTs-Au-Fc (MWCNTs: multiwalled carbon nanotubes; Fc: ferrocene) and single-strand DNA (ssDNA), semi-complementary to NoV Apt, are permanently assembled onto the SPE. By incubating Apt and methylene blue (MB), Apt-ssDNA structures with MB filling are obtained. NoV competitively combines Apt, dissociates Apt-ssDNA structures, and triggers ratiometric Δ<i>I</i>_MB/<i>I</i>_Fc. Remarkably, a programmable thermal profile is written into a micro controller unit (MCU) of the SPE. With on-chip hardware resources as the heater, error amplifier, and heater driver, the temperature curve is sectionally regulated during Apt-ssDNA regeneration, concurrently promoting reaction and preventing functionalization deterioration. Besides the wide range (1-10⁶ copies mL^-1) and impressive limit of detection (0.67 copies mL^-1), this method shows significantly less <i>I</i>_MB loss than control (98.67% vs 80.26%). For NoV-contaminated beef and pork, the proposed results meet well with certified RT-qPCR (RSDs < 4.89%).