Wang, C.Madiyar, F.Yu, C. X.Li, Jun2017-11-302017-11-302017-02-14http://hdl.handle.net/2097/38355Citation: Wang, C., Madiyar, F., Yu, C. X., & Li, J. (2017). Detection of extremely low concentration waterborne pathogen using a multiplexing self-referencing SERS microfluidic biosensor. Journal of Biological Engineering, 11, 11. doi:10.1186/s13036-017-0051-xBackground: 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.Attribution 4.0 International (CC BY 4.0)https://creativecommons.org/licenses/by/4.0/Enhanced Raman-SpectroscopyLabel-Free SersBacteriaDielectrophoresisMicroorganismsIdentificationArticle