Proteomic assay for rapid characterization of<i>Staphylococcus aureus</i>antimicrobial resistance directly from blood cultures
Abstract An efficient management of bloodstream infections requires a fast identification of the pathogen and a determination of its antimicrobial resistance profile.Staphylococcus aureusis among the most common pathogen causing bloodstream infection. A prompt characterization of methicillin-resistantStaphylococcus aureus(MRSA) and their aminoglycoside resistance profile is therefore crucial to quickly adapt the treatment and improve clinical outcomes. Among analytical technologies, targeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) has emerged as a promising tool to detect resistance mechanisms in clinical samples. Herein we present a rapid proteomic workflow to detect and quantify the most clinically relevant antimicrobial resistance effectors inS. aureus: PBP2a, PBP2c, APH(3’)-III, ANT(4’)-I, and AAC(6’)-APH(2’’), directly from positive blood cultures and in less than 70 minutes. This approach provided 99% sensitivity for PBP2a (n=98/99 strains) detection. Sensitivity was 100% for PBP2c (n=5/5), APH(3’)-III (n=16/16) and ANT(4’)-I (n=20/20), and 94% for AAC(6’)-APH(2’’) (n=16/17). Across the entire collection, 100% specificity was reported for each of the 5 resistance proteins. Additionally, relative quantification of ANT(4’)-I expression allowed to discriminate kanamycin-susceptible and -resistant strains, in strains all harboring theant(4’)-Iagene. The LC-MS/MS method presented herein demonstrates its ability to provide a reliable and in-depth profiling ofS. aureusresistance, directly from positive blood culture and in a short turnaround time, as required in clinical laboratories..
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Preprint| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
bioRxiv.org - (2023) vom: 16. Nov. Zur Gesamtaufnahme - year:2023 |
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| Sprache: |
Englisch |
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| Beteiligte Personen: |
Deforet, Francis [VerfasserIn] |
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| Links: |
Volltext [kostenfrei] |
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| doi: |
10.1101/2023.11.06.565783 |
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| PPN (Katalog-ID): |
XBI041502434 |
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| 520 | |a Abstract An efficient management of bloodstream infections requires a fast identification of the pathogen and a determination of its antimicrobial resistance profile.Staphylococcus aureusis among the most common pathogen causing bloodstream infection. A prompt characterization of methicillin-resistantStaphylococcus aureus(MRSA) and their aminoglycoside resistance profile is therefore crucial to quickly adapt the treatment and improve clinical outcomes. Among analytical technologies, targeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) has emerged as a promising tool to detect resistance mechanisms in clinical samples. Herein we present a rapid proteomic workflow to detect and quantify the most clinically relevant antimicrobial resistance effectors inS. aureus: PBP2a, PBP2c, APH(3’)-III, ANT(4’)-I, and AAC(6’)-APH(2’’), directly from positive blood cultures and in less than 70 minutes. This approach provided 99% sensitivity for PBP2a (n=98/99 strains) detection. Sensitivity was 100% for PBP2c (n=5/5), APH(3’)-III (n=16/16) and ANT(4’)-I (n=20/20), and 94% for AAC(6’)-APH(2’’) (n=16/17). Across the entire collection, 100% specificity was reported for each of the 5 resistance proteins. Additionally, relative quantification of ANT(4’)-I expression allowed to discriminate kanamycin-susceptible and -resistant strains, in strains all harboring theant(4’)-Iagene. The LC-MS/MS method presented herein demonstrates its ability to provide a reliable and in-depth profiling ofS. aureusresistance, directly from positive blood culture and in a short turnaround time, as required in clinical laboratories. | ||
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