Synthetic biology-driven optoelectronic biosensor for rapid and highly sensitive norovirus detection in fecal samples

The convergence of synthetic biology and biosensing technologies is revolutionizing diagnostics by enabling highly specific, scalable, and high-throughput detection platforms. In this study, we introduce a synthetic biology-driven optoelectronic biosensor for the rapid and highly sensitive quantification of human norovirus in clinical fecal samples. Our system integrates norovirus-like particles (NoV-LPs), produced via a baculovirus-insect cell expression system, as precise reference nanomaterials, alongside high-affinity single-chain variable fragment (scFv) antibodies generated through phage display as bioreceptors. This novel biosensing platform, implemented on a disc-based system, allows for multiplexed analysis of up to eight specimens within 45 min, eliminating the need for nucleic acid amplification. The sensor provides direct Ct-equivalent values, achieving an exceptional viral detection limit of 32 Ct-equivalent (0.5 fM), demonstrating a 10- to 100-fold improvement in sensitivity over conventional colorimetric biosensors. Clinical validation with 20 human fecal samples confirmed 100 % sensitivity and selectivity, exhibiting a strong correlation with RT-qPCR results (r = 0.945) and a relative error of 11 %. Beyond establishing a highly efficient biosensing platform, this work highlights the transformative impact of synthetic biology to engineer and produce high-affinity bioreceptors for next-generation biosensors with enhanced sensitivity, selectivity, and adaptability across diverse analytical applications.