Rapid antimicrobial susceptibility testing from positive blood cultures based on Stimulated Raman Scattering Imaging analysis

Abstract

The existing identification (ID) and antimicrobial susceptibility testing (AST) method requires at least two to three days to detect blood infection, and a fast and accurate detection method is very necessary for sepsis patients or Intensive Care Unit (ICU) patients. Here, we describe a direct isolated bacteria from a positive blood culture bottle (PBCB), and rapid AST method by femtosecond stimulated Raman scattering (SRS) imaging of deuterium oxide (D2O) metabolism, which can determine the antimicrobial susceptibility of bacteria from PBCB in 5-6 hours. The positive blood culture sample is passed through a filter membrane and mixed with cell lysis, after through the centrifugal which can directly isolated bacterium in order to identification by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS), following by antibiotic susceptibility testing by SRS imaging within a day. Overall, this rapid and rapid process combination of MALDI-TOF MS and SRS imaging of deuterium oxide (D2O) metabolism can solve the direct identification and antibiotic susceptibility testing of pathogen in positive blood cultures.