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Identification of Rare Pathogenic Bacteria in a Clinical Microbiology Laboratory by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry (CAT#: STEM-ST-0309-LJX)

Introduction

Bacterial identification directly from colonies and samples using matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry (MS) has been described as a revolutionary tool perfectly adapted to the clinical microbiology laboratory. MALDI-TOF MS has been used to identify bacterial species and subspecies, and in some outbreaks, MALDI-TOF MS has been reported to be able to identify the lineages of strains. Recently, MALDI-TOF MS has also been used to detect clinical pathogens previously misidentified or ambiguously identified. Detection of antimicrobial resistance using MALDI-TOF MS has been reported for Staphylococcus aureus, Acinetobacter baumannii, Escherichia coli, and other members of the family Enterobacteriaceae. Several new bacterial species emerging as human pathogens have been identified using MALDI-TOF MS.




Principle

In a very small area and a very short time interval (ns order of magnitude), the laser delivers high-intensity pulse energy to the sample under test, causing it to desorption and ionize instantaneously without thermal decomposition. MALDI is a mass spectrometry ionization method for direct evaporation and ionization of non-volatile samples.

Applications

For measuring the molecular weight of biological macromolecules, such as the molecular weight distribution of peptides, proteins, nucleic acids, polymers and oligomer analysis.

Procedure

1. Mix the sample with the appropriate matrix material and load it onto the metal plate.
2. Pulsed laser light is used to irradiate the sample and trigger ablation and desorption of the sample and matrix materials.
3. Analyte molecules are ionized by protonation or deprotonation in the thermal plume of the ablated gas and are then accelerated to a mass analyzer for analysis.

Materials

• Sample Type:
Rare pathogenic bacteria

Notes

1. During shutdown, if the nitrogen is not turned off, the pressure should be properly lowered to avoid moisture.
2. Keep an eye on instrument drift during manual measurement. If there is drift, the instrument needs to be calibrated with standard peptide or standard protein.