Abstract

Rapid identification and characterisation of drug-resistant bacterial pathogens have an important role in diagnostic and antimicrobial stewardship. Response time in the diagnosis of not only the etiological agent but also in antimicrobial susceptibility results is of utmost importance in patient treatment. In this study, matrix-assisted laser desorption ionisation–time of flight (MALDI-TOF) mass spectrometry (MS) was used to screen for biomarkers of ESCAPE (vancomycin-resistant Enterococcus faecium, methicillin-resistant Staphylococcus aureus, hypervirulent NAP1/ribotype 027 Clostridioides [Clostridium] difficile, multidrug resistant Acinetobacter baumannii, multidrug resistant Pseudomonas aeruginosa, and carbapenem-resistant Enterobacteriaceae) pathogens to predict antimicrobial resistance or hypervirulence. Several biomarkers of drug-resistant genotypes in S. aureus, A. baumannii, P. aeruginosa, and K. pneumoniae, as well as hypervirulence in C. difficile, were detected. The fastest possible susceptibility testing with MALDI-TOF MS is simultaneous detection of a characteristic drug-resistant peak and species identification in the same spectra generated in routine processing. According to our approach, resistance or virulence biomarker peaks can be identified while performing routine microbiology analysis, and no additional assays nor prolonged incubation time is needed. Outstanding biomarker peaks detected in our study should be further analysed by additional methods to identify the specific proteins involved.

Highlights

  • The emergence of drug-resistant bacterial strains has been accelerated by both antibiotic overuse and reduced infection control

  • MALDI-TOF mass spectrometry (MS) is widely used in clinical microbiology for bacterial identification, taxonomy, and strain typing[17]

  • Several approaches have been proposed in MALDI-TOF MS to detect antimicrobial resistance such as the detection of the entire cell profile, enzymatic activity by antibiotic hydrolysis, or resistance proteins within the cell[18,19,20,21]

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Summary

Introduction

The emergence of drug-resistant bacterial strains has been accelerated by both antibiotic overuse and reduced infection control. In 2013, the Centers for Disease Control and Prevention issued a report with the most concerning drug-resistant threats in the United States[1] Of those mentioned, the most notable are pathogens predominately associated with healthcare-associated infections (HAI)[2]. Designated by the acronym ESKAPE, this group comprises the following pathogens: vancomycin-resistant Enterococcus faecium (VREfm), methicillin-resistant Staphylococcus aureus (MRSA), carbapenem-resistant Klebsiella pneumoniae, multidrug resistant (MDR) Acinetobacter baumannii, MDR Pseudomonas aeruginosa, and carbapenem-resistant Enterobacter cloacae[3,4]. These pathogens are of considerable concern due to their high frequency in HAIs5,6. This study aimed to screen for biomarkers using MALDI-TOF MS in ESCAPE pathogens to predict antimicrobial resistance or hypervirulence

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