Detection of extremely low concentration waterborne pathogen using a multiplexing self-referencing SERS microfluidic biosensor

Background: It is challenging to achieve ultrasensitive and selective detection of waterborne pathogens at extremely low levels (i.e., single cell/mL) using conventional methods. Even with molecular methods such as ELISA or PCR, multi-enrichment steps are needed which are labor and cost intensive. In this study, we incorporated nano-dielectrophoretic microfluidic device with Surface enhanced Raman scattering (SERS) technique to build a novel portable biosensor for easy detection and characterization of Escherichia coli O157:H7 at high sensitivity level (single cell/mL). Results: A multiplexing dual recognition SERS scheme was developed to achieve one-step target detection without the need to separate target-bound probes from unbound ones. With three different SERS-tagged molecular probes targeting different epitopes of the same pathogen being deployed simultaneously, detection of pathogen targets was achieved at single cell level with sub-species specificity that has not been reported before in single-step pathogen detection. Conclusion: The self-referencing protocol implements with a Nano-dielectrophoretic microfluidic device potentially can become an easy-to-use, field-deployable spectroscopic sensor for onsite detection of pathogenic microorganisms.