Python-assisted detection and photothermal inactivation of Salmonella typhimurium and Staphylococcus aureus on a background-free SERS chip.

In this study, a background-free surface-enhanced Raman scattering (SERS) chip with a sandwich configuration was fabricated to enable reliable detection and photothermal inactivation of multiple bacteria. The SERS chip consists of a graphene-coated, phenylboronic-modified plasmonic gold substrate (pAu/G/PBA), and two aptamer-functionalized core (gold)-shell (Prussian blue/Poly-L-lysine and 4-mercaptobenzonitrile/polydopamine) SERS tags (Au@PB@PLL@Apt and Au@MB@PDA@Apt). The detection signals rely on the characteristic and nonoverlapping Raman bands of the SERS tags within the Raman-silent region (1800-2800 cm-1 ), where no background signals from the sample matrix are observed, leading to improved detection sensitivity and accuracy. Considering the relatively large size of bacteria (e.g., micron level), a rapid Raman mapping technique was chosen over conventional point-scan methods to achieve more reliable quantitative analysis of bacteria. This technique involves collecting and analyzing intensity signals of SERS tags from all the scattering points with an average ensemble effect, which is facilitated by the use of Python. As a proof-of-concept, model bacterium of Salmonella typhimurium and Staphylococcus aureus were successfully detected using the SERS chip with a dynamic range of 10-107  CFU/mL. Additionally, the SERS chip demonstrated successful detection of these bacteria in whole blood samples. Moreover, the photothermal effect of pAu/G led to efficient bacteria elimination, achieving approximately 100% eradication. This study integrated a background-free SERS chip with a Python-assisted rapid Raman mapping technique, resulting in a reliable, rapid and accurate method for detecting and eliminating multiple bacteria, which may provide a promising alternative for multiple screening of bacteria in real samples.