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Rapid MRSA detection via tandem mass spectrometry of the intact 80 kDa PBP2a resistance protein

Treatment of antibiotic-resistant infections is dependent on the detection of specific bacterial genes or proteins in clinical assays. Identification of methicillin-resistant Staphylococcus aureus (MRSA) is often accomplished through the detection of penicillin-binding protein 2a (PBP2a). With great...

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Autores principales: Neil, Jason R., Verma, Arvind, Kronewitter, Scott R., McGee, William M., Mullen, Christopher, Viirtola, Marjaana, Kotovuori, Annika, Friedrich, Herdis, Finell, Johan, Rannisto, Joni, Syka, John E. P., Stephenson, James L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8443585/
https://www.ncbi.nlm.nih.gov/pubmed/34526615
http://dx.doi.org/10.1038/s41598-021-97844-w
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author Neil, Jason R.
Verma, Arvind
Kronewitter, Scott R.
McGee, William M.
Mullen, Christopher
Viirtola, Marjaana
Kotovuori, Annika
Friedrich, Herdis
Finell, Johan
Rannisto, Joni
Syka, John E. P.
Stephenson, James L.
author_facet Neil, Jason R.
Verma, Arvind
Kronewitter, Scott R.
McGee, William M.
Mullen, Christopher
Viirtola, Marjaana
Kotovuori, Annika
Friedrich, Herdis
Finell, Johan
Rannisto, Joni
Syka, John E. P.
Stephenson, James L.
author_sort Neil, Jason R.
collection PubMed
description Treatment of antibiotic-resistant infections is dependent on the detection of specific bacterial genes or proteins in clinical assays. Identification of methicillin-resistant Staphylococcus aureus (MRSA) is often accomplished through the detection of penicillin-binding protein 2a (PBP2a). With greater dependence on mass spectrometry (MS)-based bacterial identification, complementary efforts to detect resistance have been hindered by the complexity of those proteins responsible. Initial characterization of PBP2a indicates the presence of glycan modifications. To simplify detection, we demonstrate a proof-of-concept tandem MS approach involving the generation of N-terminal PBP2a peptide-like fragments and detection of unique product ions during top-down proteomic sample analyses. This approach was implemented for two PBP2a variants, PBP2a(mecA) and PBP2a(mecC), and was accurate across a representative panel of MRSA strains with different genetic backgrounds. Additionally, PBP2a(mecA) was successfully detected from clinical isolates using a five-minute liquid chromatographic separation and implementation of this MS detection strategy. Our results highlight the capability of direct MS-based resistance marker detection and potential advantages for implementing these approaches in clinical diagnostics.
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spelling pubmed-84435852021-09-20 Rapid MRSA detection via tandem mass spectrometry of the intact 80 kDa PBP2a resistance protein Neil, Jason R. Verma, Arvind Kronewitter, Scott R. McGee, William M. Mullen, Christopher Viirtola, Marjaana Kotovuori, Annika Friedrich, Herdis Finell, Johan Rannisto, Joni Syka, John E. P. Stephenson, James L. Sci Rep Article Treatment of antibiotic-resistant infections is dependent on the detection of specific bacterial genes or proteins in clinical assays. Identification of methicillin-resistant Staphylococcus aureus (MRSA) is often accomplished through the detection of penicillin-binding protein 2a (PBP2a). With greater dependence on mass spectrometry (MS)-based bacterial identification, complementary efforts to detect resistance have been hindered by the complexity of those proteins responsible. Initial characterization of PBP2a indicates the presence of glycan modifications. To simplify detection, we demonstrate a proof-of-concept tandem MS approach involving the generation of N-terminal PBP2a peptide-like fragments and detection of unique product ions during top-down proteomic sample analyses. This approach was implemented for two PBP2a variants, PBP2a(mecA) and PBP2a(mecC), and was accurate across a representative panel of MRSA strains with different genetic backgrounds. Additionally, PBP2a(mecA) was successfully detected from clinical isolates using a five-minute liquid chromatographic separation and implementation of this MS detection strategy. Our results highlight the capability of direct MS-based resistance marker detection and potential advantages for implementing these approaches in clinical diagnostics. Nature Publishing Group UK 2021-09-15 /pmc/articles/PMC8443585/ /pubmed/34526615 http://dx.doi.org/10.1038/s41598-021-97844-w Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Neil, Jason R.
Verma, Arvind
Kronewitter, Scott R.
McGee, William M.
Mullen, Christopher
Viirtola, Marjaana
Kotovuori, Annika
Friedrich, Herdis
Finell, Johan
Rannisto, Joni
Syka, John E. P.
Stephenson, James L.
Rapid MRSA detection via tandem mass spectrometry of the intact 80 kDa PBP2a resistance protein
title Rapid MRSA detection via tandem mass spectrometry of the intact 80 kDa PBP2a resistance protein
title_full Rapid MRSA detection via tandem mass spectrometry of the intact 80 kDa PBP2a resistance protein
title_fullStr Rapid MRSA detection via tandem mass spectrometry of the intact 80 kDa PBP2a resistance protein
title_full_unstemmed Rapid MRSA detection via tandem mass spectrometry of the intact 80 kDa PBP2a resistance protein
title_short Rapid MRSA detection via tandem mass spectrometry of the intact 80 kDa PBP2a resistance protein
title_sort rapid mrsa detection via tandem mass spectrometry of the intact 80 kda pbp2a resistance protein
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8443585/
https://www.ncbi.nlm.nih.gov/pubmed/34526615
http://dx.doi.org/10.1038/s41598-021-97844-w
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