This project aims to develop a versatile and easy-to-use surface plasmon resonance imaging (SPRi) instrument for the rapid and broad-spectrum detection of low concentrations of pathogenic bacteria in complex samples, particularly blood. SPRi is a label-free technique that allows real-time probing of a sample (regardless of its optical transparency). Due to the high sensitivity of the plasmon phenomenon, the dynamic range of measurable index variation is limited by SPRi detection when reading is performed at a fixed angle, as is the case in commercially available devices. This reduces the use of such optical instruments to the study of environments whose index remains relatively stable during the experiment and whose molecular probes have molecular weights comparable to the targets (monitoring of bimolecular interactions).
This considerably limits the detection of growing bacteria in complex environments. Our laboratory has developed original solutions for the detection of very low levels of contamination in food matrices (creation of a start-up in 2012), but SPRi is unsuitable for the detection of bacteria in blood, partly due to the very high intrinsic variability of this matrix.
These limitations will be overcome by integrating five complementary components:
1. The design of an instrument optimized for real-time recording of SPR images over a defined range of illumination angles;
2. The development of dedicated SPR data analysis and processing to extract the most relevant information for each probe from the images in real time;
3. The functionalization of biochips through a combination of appropriate probes (series of peptides such as antimicrobial peptides (AMPs), antibodies, and even bacteriophages) to optimize the number of possible identifications with a reduced set of probes;
4. The learning of specific “4D-SPRi signatures” of model strains in blood matrices;
5. Validation of the performance of the new “4D-SPRi” instrument as a tool for detecting and characterizing bacteria from hospital strains compared to reference techniques.