Assessment of bacterial viability by laser desorption ionization mass spectrometry for antimicrobial susceptibility testing

Abstract

Bacterial infection poses a serious threat to human health worldwide. Rapid antimicrobial susceptibility testing (AST) is essential for the clinical treatment of bacterial infection patients. However, the traditional AST relies on bacteria culture, which is time-consuming and limits the analysis to culturable species. Herein, we present a laser desorption ionization (LDI) mass spectrometry-based method for rapid bacterial viability assessment and AST by tracing the redox of resazurin (RS) by viable bacteria. RS as well as its reduction product, fluorescent resorufin (RF), can be directly detected by LDI-MS in the absence of matrix. The intensity ratio between RF and RS can be used to assess the viability of bacteria in specimens. We have demonstrated the high efficiency of the method using different bacterial species, including K. pneumoniae, S. aureus, E. coli, and P. aeruginosa, and various antibiotic drugs, such as ciprofloxacin, ampicillin, tetracycline, oxytetracycline, ciprofloxacin and levofloxacin. Compared to traditional methods based on optical absorption, the current method is faster and more sensitive. Furthermore, we applied the method to bacterial viability detection and AST using human body fluid samples, i.e. serum and urine, demonstrating that it can screen rapidly appropriate antibiotic drugs for timely clinical treatment of infectious diseases. With the advantages of simplicity in methodology as well as sensitivity and speed in analysis, the current method holds the potential of clinical usages.