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, 9 (1), 18945

Screening of Biomarkers of Drug Resistance or Virulence in ESCAPE Pathogens by MALDI-TOF Mass Spectrometry

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Screening of Biomarkers of Drug Resistance or Virulence in ESCAPE Pathogens by MALDI-TOF Mass Spectrometry

Samantha Flores-Treviño et al. Sci Rep.

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.

Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
Representative mass spectra of peak m/z 4,594 in S. aureus. The peak m/z 4,594 is presented as a doublet in MRSA isolates (red line) and as a singlet in MSSA isolates (green line). The average spectrum of each group of peak m/z 4,594 (a) and the spectra of all the analysed isolates (b) are shown.
Figure 2
Figure 2
Representative mass spectra of peaks m/z 6,654 and 6,712 in C. difficile. The peak m/z 6,654 is present in non-027 ribotype isolates (green line) and is absent in 027 and 176 ribotype (red line). By contrast, the peak m/z 6,712 is present in 027 and ribotypes but absent in non-027 ribotypes. The average spectrum of each group of peaks m/z 6,654 and 6,712 (a) and the spectra of all the analysed isolates (b) are shown.
Figure 3
Figure 3
Representative mass spectra of peaks m/z 6,304 and 6,332 in A. baumannii. The peak m/z 6,304 is present in blaOXA-58 isolates (green line) and is absent in blaOXA-24 isolates (red line). By contrast, the peak m/z 6,332 is present in blaOXA-24 but absent in blaOXA-58 isolates. The average spectrum of each group of peaks m/z 6,304 and 6,332 (a) and the spectra of all the analysed isolates (b) are shown.
Figure 4
Figure 4
Representative mass spectra of peaks m/z 2,726, 5,455 and 5,742 in P. aeruginosa. Three peaks m/z 2,726, 5,455, and 5,742 were present in the majority of non-MDR isolates (green line) and at higher intensities than those in MDR isolates (red line). The average spectrum of each group of peaks m/z 2,726 (a), 5,455 (c), and 5,742 (e) and the spectra of all the analysed isolates for peaks m/z 2,726 (b), 5,455 (d), and 5,742 (f) are shown.
Figure 5
Figure 5
Representative mass spectra of peaks m/z 6,100 in K. pneumoniae. The peak m/z 6,100 is absent in the carbapenem- and colistin-resistant isolates (red line) and is present in the majority of the carbapenem- and colistin-susceptible isolates (green line). The average spectrum of each group of peak m/z 6,100 (a) and the spectra of all the analysed isolates (b) are shown.

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