Rapid Detection of Salmonella typhimurium in Drinking Water by a White Light Reflectance Spectroscopy Immunosensor

Biosensors represent an attractive approach for fast bacteria detection. Here, we present an optical biosensor for the detection of Salmonella typhimurium lipopolysaccharide (LPS) and Salmonella bacteria in drinking water, based on white light reflectance spectroscopy. The sensor chip consisted of a Si die with a thin SiO₂ layer on top that was transformed into a biosensor through the immobilization of Salmonella LPS. The optical setup included a reflection probe with seven 200 μm fibers, a visible and near-infrared light source, and a spectrometer. The six fibers at the reflection probe circumference were coupled with the light source and illuminated the biosensor chip vertically, whereas the central fiber collected the reflected light and guided it to the spectrometer. A competitive immunoassay configuration was adopted for the analysis. Accordingly, a mixture of LPS or bacteria solution, pre-incubated for 15 min, with an anti-Salmonella LPS antibody was pumped over the chip followed by biotinylated secondary antibody and streptavidin for signal enhancement. The binding of the free anti-Salmonella antibody to chip-immobilized LPS led to a shift of the reflectance spectrum that was inversely related to the analyte concentration (LPS or bacteria) in the calibrators or samples. The total assay duration was 15 min, and the detection limits achieved were 4 ng/mL for LPS and 320 CFU/mL for bacteria. Taking into account the low detection limits, the short analysis time, and the small size of the chip and instrumentation employed, the proposed immunosensor could find wide application for bacteria detection in drinking water.