Flash MS/MS proteotyping allows identifying microbial isolates in 36 s of mass spectrometry signal

Area of Science:

• Microbiology
• Analytical Chemistry
• Bioinformatics

Background:

• Rapid microbial identification is crucial for diagnostics, sanitation, and food safety.
• Current whole-cell MALDI-TOF mass spectrometry has limitations in database completeness and discrimination power, often restricted to species level.
• Environmental isolates and opportunistic pathogens pose challenges for existing methods.

Purpose of the Study:

• To develop an ultra-fast microbial identification methodology using tandem mass spectrometry.
• To enhance taxonomic discrimination beyond the species level, down to the subspecies or strain level.
• To enable high-throughput analysis for applications like culturomics and diagnostics.

Main Methods:

• Direct sample infusion tandem mass spectrometry for ultra-fast data acquisition.
• Development of a sensitive data processing and taxonomic identification procedure.
• Validation using reference strains and uncharacterized bacterial isolates.

Main Results:

• Microbial identification to the species level achieved in 36 seconds of tandem mass spectrometry signal.
• Total analysis time, including injection, was 102 seconds.
• The 'flash proteotyping' method demonstrated high discrimination, providing information down to the strain level.

Conclusions:

• Tandem mass spectrometry-based flash proteotyping offers a rapid and highly discriminating method for microbial identification.
• This methodology overcomes limitations of current techniques, particularly for environmental and challenging isolates.
• The approach holds significant potential for advancing high-throughput microbial analysis in various fields, including diagnostics and culturomics.