Detection of stx2 from Shiga toxin-producing Escherichia coli (STEC) by a surface enhanced Raman spectroscopy (SERS) sensor using recycled silicon chips

Abstract

In this research, a selective, cost-efficient, and highly sensitive Ag nanostructure Surface Enhanced Raman Spectroscopy (SERS) sensor was developed as a methodological approach to rapidly detect a targetss-DNA (stx2) in STEC (Shiga toxin-producing Escherichia coli). The Ag nanostructure-based SERS substrate was functionalized by two types of thiols: thiol-ss-DNA for bonding target ss-DNA and 6-Mercapto-1-hexanol (HS(CH2)6OH) for blocking the Ag nanostructure surface. Methylene Blue (MB) was used as a Raman marker to quantify target ss-DNA, as well as a model molecule to characterize the electrodeposited Ag nanostructure SERS substrate. Ag nanostructure SERS substrates showed good sensitivity and repeatability towards MB detection, with a LOD = 0.3158 μM, and RSD = 12.48% (at 45 different random points for 0.1 μM MB). More importantly, the Ag nanostructure/ss-DNA SERS substrate showed good selectivity towards STEC O157 stx2 target DNA, as well as good linearity and sensitivity towards its detection in a buffer solution. A limit of detection of 0.4900 aM and a wide linear range from 1 aM to 100 pM were demonstrated. The SERS sensors were able to identify target DNA (stx2) in a STEC strain and the study showed proof of principle that SERS substrate has potential as a cost-effective, highly selective, highly sensitive DNA and bacteria sensor without the aid of DNA amplification. With further development and validation, this methodological approach has the potential for point-of-use detection for instance on a farm or in the food industry.