Multiplex Detection and Quantification of Virus Co-Infections Using Label-free Surface-Enhanced Raman Spectroscopy and Deep Learning Algorithms

Abstract

Multiple respiratory viruses can concurrently or sequentially infect the respiratory tract, making their identification crucial for diagnosis, treatment, and disease management. We present a label-free diagnostic platform integrating surface-enhanced Raman scattering (SERS) with deep learning for rapid, quantitative detection of respiratory virus coinfections. Using sensitive silica-coated silver nanorod array substrates, over 1.2 million SERS spectra are collected from 11 viruses, nine two-virus mixtures, and four three-virus mixtures at various concentrations in saliva. A deep learning model, MultiplexCR, is developed to simultaneously predict virus species and concentrations from SERS spectra. It achieves an impressive 98.6% accuracy in classifying virus coinfections and a mean absolute error of 0.028 for concentration regression. In blind tests, the model demonstrates consistent high accuracy and precise concentration predictions. This SERS-MultiplexCR platform completes the entire detection process in just 15 min, offering significant potential for rapid, point-of-care diagnostics in infection detection, as well as applications in food safety and environmental monitoring.