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Identification of Staphylococcus aureus and Determination of Its Methicillin Resistance by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (CAT#: STEM-ST-0322-LJX)

Introduction

To evaluate the performance of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) in identifying Staphylococcus aureus and in determining its methicillin resistance, we analyzed 76 S. aureus clinical isolates using a linear MALDI-TOF MS. Spectral profile data obtained were compared with the database provided with the instrument, and 74% of the isolates were identified as S. aureus, as confirmed by a nuc-based PCR test. The determination of the methicillin resistance in S. aureus is based on the fact that the spectral profiles of methicillin-susceptible S. aureus (MSSA) and methicillin-resistant S. aureus (MRSA) differ greatly from each other. Replicate spectral profiles obtained from each isolate were combined to be a representative spectrum of it, and representative spectral profiles from all the isolates constitute a user's self-established database. All the spectral profiles in the database were classified into two groups based on clustering analysis, and one is for MSSA and another MRSA. There was a little discrepancy between the results from MALDI-TOF MS and from PCR. Seven isolates that are negative for the mecA gene by PCR were identified as MRSA by 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:
Staphylococcus aureus

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.
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