Enzymatic Activity Profiling Using an Ultra-Sensitive Array of Chemiluminescent Probes for Bacterial Classification and Characterization

Bacterial species identification and characterization in clinical and industrial settings necessitate the use of diverse, labor-intensive, and time-consuming protocols, as well as the utilization of expensive and high-maintenance equipment. Furthermore, while cutting-edge identification technologies such as mass spectrometry and PCR are highly effective in identifying bacterial pathogens, they fall short in providing crucial information for identifying bacteria that are not present in the databases on which these methods rely. In response to these challenges, we present a robust and general approach to bacterial identification based on their unique enzymatic activity profiles. This method delivers results within 90 minutes, utilizing an array of highly sensitive and enzyme-selective chemiluminescent probes. Leveraging our recently developed technology of chemiluminescent luminophores, which emit light under physiological conditions, we have crafted an array of probes designed to rapidly detect various bacterial hydrolytic enzymatic activities, including some associated with antibiotic resistance. The analysis of chemiluminescent fingerprints from a diverse panel of prominent bacterial pathogens has revealed distinct enzymatic activity profiles for each strain. The reported universally applicable identification procedure offers a highly sensitive and expeditious means to delineate bacterial enzymatic activity profiles. This breakthrough opens new avenues for characterizing and identifying pathogens in research, clinical, and industrial applications.