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Methodology for the Study of Horizontal Gene Transfer in Staphylococcus aureus

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Different Vancomycin-Intermediate Staphylococcus aureus Phenotypes Selected from the Same ST100-hVISA Parental

Sabrina Di Gregorio¹, Silvina Fernandez¹, Arabela Cuirolo²

¹1 Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Departamento de Microbiología, Inmunología y Biotecnología, Cátedra de Microbiología, Buenos Aires, Argentina .

Microbial Drug Resistance (Larchmont, N.Y.)

|December 20, 2016

Summary

This study characterizes peptidoglycan metabolism in vancomycin-intermediate Staphylococcus aureus (VISA) ST100 strains. Different VISA phenotypes emerged from a single genetic background, impacting cell wall structure and antimicrobial susceptibility.

Keywords:

MRSA ST100 Staphylococcus aureus VISA hVISA vancomycin

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Area of Science:

Microbiology
Molecular Biology
Genetics

Background:

The emergence of vancomycin-intermediate Staphylococcus aureus (VISA) poses a significant clinical challenge.
Understanding the genetic and metabolic factors driving VISA development is crucial for effective treatment strategies.

Purpose of the Study:

To investigate the factors influencing peptidoglycan metabolism in isogenic hVISA/VISA ST100 strains.
To characterize the differences between clinical and laboratory-selected VISA strains and their mutants.

Main Methods:

High-performance liquid chromatography-mass spectrometry (HPLC-MS) for peptidoglycan muropeptide analysis.
Analysis of cell wall thickness and gene expression (pbp2, pbp4).
Assessment of autolysis profiles and antimicrobial susceptibility.

Main Results:

Vancomycin treatment and selection led to increased monomeric and decreased tetrameric muropeptides in VISA strains.
VISA strains exhibited increased cell wall thickness and elevated pbp2 gene expression.
Distinct VISA phenotypes emerged with variations in autolysis and pbp4 transcription, affecting antimicrobial susceptibility.

Conclusions:

A single hVISA ST100 genetic background can give rise to diverse VISA phenotypes.
Alterations in peptidoglycan metabolism and cell wall synthesis are key mechanisms in VISA development.
These findings have implications for understanding VISA evolution and treatment resistance.