Mass Spectrometry Proteotyping-Based Detection and Identification of Staphylococcus aureus, Escherichia coli, and Candida albicans in Blood
Bloodstream infections (BSIs), the presence of microorganisms in blood, are potentially serious conditions that can quickly develop into sepsis and life-threatening situations. When assessing proper treatment, rapid diagnosis is the key; besides clinical judgement performed by attending physicians,...
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Frontiers Media S.A.
2021-07-01
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Series: | Frontiers in Cellular and Infection Microbiology |
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Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2021.634215/full |
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Article |
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DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Nahid Kondori Nahid Kondori Amra Kurtovic Beatriz Piñeiro-Iglesias Francisco Salvà-Serra Francisco Salvà-Serra Francisco Salvà-Serra Francisco Salvà-Serra Daniel Jaén-Luchoro Daniel Jaén-Luchoro Björn Andersson Gelio Alves Aleksey Ogurtsov Annika Thorsell Johannes Fuchs Timur Tunovic Nina Kamenska Anders Karlsson Yi-Kuo Yu Edward R. B. Moore Edward R. B. Moore Edward R. B. Moore Roger Karlsson Roger Karlsson Roger Karlsson |
spellingShingle |
Nahid Kondori Nahid Kondori Amra Kurtovic Beatriz Piñeiro-Iglesias Francisco Salvà-Serra Francisco Salvà-Serra Francisco Salvà-Serra Francisco Salvà-Serra Daniel Jaén-Luchoro Daniel Jaén-Luchoro Björn Andersson Gelio Alves Aleksey Ogurtsov Annika Thorsell Johannes Fuchs Timur Tunovic Nina Kamenska Anders Karlsson Yi-Kuo Yu Edward R. B. Moore Edward R. B. Moore Edward R. B. Moore Roger Karlsson Roger Karlsson Roger Karlsson Mass Spectrometry Proteotyping-Based Detection and Identification of Staphylococcus aureus, Escherichia coli, and Candida albicans in Blood Frontiers in Cellular and Infection Microbiology blood-stream infections proteotyping MALDI-TOF MS proteomics bacteremia fungemia |
author_facet |
Nahid Kondori Nahid Kondori Amra Kurtovic Beatriz Piñeiro-Iglesias Francisco Salvà-Serra Francisco Salvà-Serra Francisco Salvà-Serra Francisco Salvà-Serra Daniel Jaén-Luchoro Daniel Jaén-Luchoro Björn Andersson Gelio Alves Aleksey Ogurtsov Annika Thorsell Johannes Fuchs Timur Tunovic Nina Kamenska Anders Karlsson Yi-Kuo Yu Edward R. B. Moore Edward R. B. Moore Edward R. B. Moore Roger Karlsson Roger Karlsson Roger Karlsson |
author_sort |
Nahid Kondori |
title |
Mass Spectrometry Proteotyping-Based Detection and Identification of Staphylococcus aureus, Escherichia coli, and Candida albicans in Blood |
title_short |
Mass Spectrometry Proteotyping-Based Detection and Identification of Staphylococcus aureus, Escherichia coli, and Candida albicans in Blood |
title_full |
Mass Spectrometry Proteotyping-Based Detection and Identification of Staphylococcus aureus, Escherichia coli, and Candida albicans in Blood |
title_fullStr |
Mass Spectrometry Proteotyping-Based Detection and Identification of Staphylococcus aureus, Escherichia coli, and Candida albicans in Blood |
title_full_unstemmed |
Mass Spectrometry Proteotyping-Based Detection and Identification of Staphylococcus aureus, Escherichia coli, and Candida albicans in Blood |
title_sort |
mass spectrometry proteotyping-based detection and identification of staphylococcus aureus, escherichia coli, and candida albicans in blood |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Cellular and Infection Microbiology |
issn |
2235-2988 |
publishDate |
2021-07-01 |
description |
Bloodstream infections (BSIs), the presence of microorganisms in blood, are potentially serious conditions that can quickly develop into sepsis and life-threatening situations. When assessing proper treatment, rapid diagnosis is the key; besides clinical judgement performed by attending physicians, supporting microbiological tests typically are performed, often requiring microbial isolation and culturing steps, which increases the time required for confirming positive cases of BSI. The additional waiting time forces physicians to prescribe broad-spectrum antibiotics and empirically based treatments, before determining the precise cause of the disease. Thus, alternative and more rapid cultivation-independent methods are needed to improve clinical diagnostics, supporting prompt and accurate treatment and reducing the development of antibiotic resistance. In this study, a culture-independent workflow for pathogen detection and identification in blood samples was developed, using peptide biomarkers and applying bottom-up proteomics analyses, i.e., so-called “proteotyping”. To demonstrate the feasibility of detection of blood infectious pathogens, using proteotyping, Escherichia coli and Staphylococcus aureus were included in the study, as the most prominent bacterial causes of bacteremia and sepsis, as well as Candida albicans, one of the most prominent causes of fungemia. Model systems including spiked negative blood samples, as well as positive blood cultures, without further culturing steps, were investigated. Furthermore, an experiment designed to determine the incubation time needed for correct identification of the infectious pathogens in blood cultures was performed. The results for the spiked negative blood samples showed that proteotyping was 100- to 1,000-fold more sensitive, in comparison with the MALDI-TOF MS-based approach. Furthermore, in the analyses of ten positive blood cultures each of E. coli and S. aureus, both the MALDI-TOF MS-based and proteotyping approaches were successful in the identification of E. coli, although only proteotyping could identify S. aureus correctly in all samples. Compared with the MALDI-TOF MS-based approaches, shotgun proteotyping demonstrated higher sensitivity and accuracy, and required significantly shorter incubation time before detection and identification of the correct pathogen could be accomplished. |
topic |
blood-stream infections proteotyping MALDI-TOF MS proteomics bacteremia fungemia |
url |
https://www.frontiersin.org/articles/10.3389/fcimb.2021.634215/full |
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doaj-bd737f0c29ee4ee58af47563e061cd862021-07-26T12:59:41ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882021-07-011110.3389/fcimb.2021.634215634215Mass Spectrometry Proteotyping-Based Detection and Identification of Staphylococcus aureus, Escherichia coli, and Candida albicans in BloodNahid Kondori0Nahid Kondori1Amra Kurtovic2Beatriz Piñeiro-Iglesias3Francisco Salvà-Serra4Francisco Salvà-Serra5Francisco Salvà-Serra6Francisco Salvà-Serra7Daniel Jaén-Luchoro8Daniel Jaén-Luchoro9Björn Andersson10Gelio Alves11Aleksey Ogurtsov12Annika Thorsell13Johannes Fuchs14Timur Tunovic15Nina Kamenska16Anders Karlsson17Yi-Kuo Yu18Edward R. B. Moore19Edward R. B. Moore20Edward R. B. Moore21Roger Karlsson22Roger Karlsson23Roger Karlsson24Department of Infectious Diseases, Sahlgrenska Academy, University of Gothenburg, Gothenburg, SwedenDepartment of Clinical Microbiology, Sahlgrenska University Hospital, Gothenburg, SwedenDepartment of Clinical Microbiology, Sahlgrenska University Hospital, Gothenburg, SwedenDepartment of Clinical Microbiology, Sahlgrenska University Hospital, Gothenburg, SwedenDepartment of Infectious Diseases, Sahlgrenska Academy, University of Gothenburg, Gothenburg, SwedenDepartment of Clinical Microbiology, Sahlgrenska University Hospital, Gothenburg, SwedenCulture Collection University of Gothenburg (CCUG), Sahlgrenska Academy of the University of Gothenburg, Gothenburg, SwedenMicrobiology, Department of Biology, University of the Balearic Islands, Palma de Mallorca, SpainDepartment of Infectious Diseases, Sahlgrenska Academy, University of Gothenburg, Gothenburg, SwedenCulture Collection University of Gothenburg (CCUG), Sahlgrenska Academy of the University of Gothenburg, Gothenburg, SwedenBioinformatics Core Facility at Sahlgrenska Academy, University of Gothenburg, Gothenburg, SwedenNational Center for Biotechnology Information (NCBI), Bethesda, MD, United StatesNational Center for Biotechnology Information (NCBI), Bethesda, MD, United StatesProteomics Core Facility at Sahlgrenska Academy, University of Gothenburg, Gothenburg, SwedenProteomics Core Facility at Sahlgrenska Academy, University of Gothenburg, Gothenburg, SwedenDepartment of Clinical Microbiology, Sahlgrenska University Hospital, Gothenburg, SwedenNorra-Älvsborgs-Länssjukhus (NÄL), Trollhättan, SwedenNanoxis Consulting AB, Gothenburg, SwedenNational Center for Biotechnology Information (NCBI), Bethesda, MD, United StatesDepartment of Infectious Diseases, Sahlgrenska Academy, University of Gothenburg, Gothenburg, SwedenDepartment of Clinical Microbiology, Sahlgrenska University Hospital, Gothenburg, SwedenCulture Collection University of Gothenburg (CCUG), Sahlgrenska Academy of the University of Gothenburg, Gothenburg, SwedenDepartment of Infectious Diseases, Sahlgrenska Academy, University of Gothenburg, Gothenburg, SwedenDepartment of Clinical Microbiology, Sahlgrenska University Hospital, Gothenburg, SwedenNanoxis Consulting AB, Gothenburg, SwedenBloodstream infections (BSIs), the presence of microorganisms in blood, are potentially serious conditions that can quickly develop into sepsis and life-threatening situations. When assessing proper treatment, rapid diagnosis is the key; besides clinical judgement performed by attending physicians, supporting microbiological tests typically are performed, often requiring microbial isolation and culturing steps, which increases the time required for confirming positive cases of BSI. The additional waiting time forces physicians to prescribe broad-spectrum antibiotics and empirically based treatments, before determining the precise cause of the disease. Thus, alternative and more rapid cultivation-independent methods are needed to improve clinical diagnostics, supporting prompt and accurate treatment and reducing the development of antibiotic resistance. In this study, a culture-independent workflow for pathogen detection and identification in blood samples was developed, using peptide biomarkers and applying bottom-up proteomics analyses, i.e., so-called “proteotyping”. To demonstrate the feasibility of detection of blood infectious pathogens, using proteotyping, Escherichia coli and Staphylococcus aureus were included in the study, as the most prominent bacterial causes of bacteremia and sepsis, as well as Candida albicans, one of the most prominent causes of fungemia. Model systems including spiked negative blood samples, as well as positive blood cultures, without further culturing steps, were investigated. Furthermore, an experiment designed to determine the incubation time needed for correct identification of the infectious pathogens in blood cultures was performed. The results for the spiked negative blood samples showed that proteotyping was 100- to 1,000-fold more sensitive, in comparison with the MALDI-TOF MS-based approach. Furthermore, in the analyses of ten positive blood cultures each of E. coli and S. aureus, both the MALDI-TOF MS-based and proteotyping approaches were successful in the identification of E. coli, although only proteotyping could identify S. aureus correctly in all samples. Compared with the MALDI-TOF MS-based approaches, shotgun proteotyping demonstrated higher sensitivity and accuracy, and required significantly shorter incubation time before detection and identification of the correct pathogen could be accomplished.https://www.frontiersin.org/articles/10.3389/fcimb.2021.634215/fullblood-stream infectionsproteotypingMALDI-TOF MSproteomicsbacteremiafungemia |