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MALDI-TOF Mass Spectrometry: Principle and Applications in Microbiology
MALDI-TOF Mass Spectrometry: Principle and Applications in Microbiology
MALDI stands for Matrix-Assisted Laser Desorption Ionization. In this ionization method
samples are fixed in a crystalline matrix and are bombarded by a laser. The sample molecules
vaporize into the vacuum while being ionized at the same time without fragmenting or
decomposing.
TOF stands for Time of Flight, a mass spectrometry method that separates ions by their mass to
charge ratio and determines that mass to charge ratio by the time it takes for the ions to reach a
detector.
This technology generates characteristic mass spectral fingerprints which is compared with large
library of mass spectra. As the spectral fingerprints are unique signatures for each microorganism
accurate microbial identification at the genus and species levels is done using bioinformatics
pattern profiling.
1. Ionization Phase
2. Time of Flight Phase
Ionization Phase:
Initially, the samples are fixed in a crystalline matrix in a target plate and are bombarded by a
laser. The sample molecules vaporize into the vacuum while being ionized at the same time.
High voltage is then applied to accelerate the charged particles.
1. In the linear mode, particles will impinge upon the linear detector within a few
nanoseconds after ionization. Higher mass molecules will arrive later than lighter ones.
Flight time measurement make it possible to determine molecule masses directly. Each
peak in the spectrum corresponds to the specific mass of the particle along the time axis,
starting with the ionization moment.
2. In the reflector mode, the particles are diverted so that they fly towards a second
detector. In addition to extending the flight distance, the reflector also focuses the masses.
The combination of these two effects makes for higher resolution than in the linear mode.
The net result is a generation of a mass spectrum which is compared with those of well-
characterized organisms available in the reference library database to identify the isolate.
Procedure
Typical workflow
Applications in Microbiology
Microbial identification by MALDI-TOF MS has skyrocketed over the last couple of years
because it offers species-level identifications in minutes at low costs with accuracy that matches
and often exceeds that of conventional identification systems.
MALDI-TOF MS is being used for routine diagnostic or diagnostic-like purposes in clinic,
veterinary, pharma and food microbiology (food quality control) laboratories as well as for
environmental monitoring, biodefense and various biological research.
Limitations
Identification of new isolates is possible only if the spectral database contains peptide
mass fingerprints of the type strains of specific genera/species/subspecies/strains
No susceptibility information is provided
Not useful for direct testing of clinical specimens (except urine)
Some organisms require repeat analysis and additional processing (extraction)
The acceptable score cutoffs vary between studies and some closely related organisms are
not differentiated.
Some organisms currently cannot be reliably identified by this method, such
as Shigella spp and Streptococcus pneumoniae.