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Influence of antibiotic treatment on the detection of S. aureus in whole blood following pathogen enrichment
BACKGROUND: Early pathogen detection and identification are crucial for an effective and targeted antibiotic therapy in patients suffering from blood stream infection. Molecular diagnostic methods can accelerate pathogen identification as compared to blood culture, but frequently suffer from the inh...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6683330/ https://www.ncbi.nlm.nih.gov/pubmed/31387527 http://dx.doi.org/10.1186/s12866-019-1559-7 |
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author | Pilecky, Matthias Schildberger, Anita Knabl, Ludwig Orth-Höller, Dorothea Weber, Viktoria |
author_facet | Pilecky, Matthias Schildberger, Anita Knabl, Ludwig Orth-Höller, Dorothea Weber, Viktoria |
author_sort | Pilecky, Matthias |
collection | PubMed |
description | BACKGROUND: Early pathogen detection and identification are crucial for an effective and targeted antibiotic therapy in patients suffering from blood stream infection. Molecular diagnostic methods can accelerate pathogen identification as compared to blood culture, but frequently suffer from the inhibition of polymerase chain reation (PCR) by sample matrix components, such as host DNA, anticoagulants, or plasma proteins. To overcome this limitation, molecular diagnostic methods commonly rely on pathogen enrichment by selective lysis of blood cells and pelleting of intact pathogens prior to analysis. RESULTS: Here, we investigated the impact of antibiotic treatment on the recovery of pathogen DNA using an established pathogen enrichment protocol. Based on the hypothesis that induction of bacterial cell wall disintegration following antibiotic administration leads to incomplete pelleting of pathogen DNA, S. aureus was grown in human whole blood with or without addition of cell wall active (vancomycin, piperacillin) or non cell wall active (ciprofloxacin, clindamycin) antibiotics at clinically relevant concentrations. Pathogen detection remained unaffected by non cell wall active antibiotics or even increased in the presence of cell wall active antibiotics, indicating improved accessibility of pathogen DNA. Likewise, mechanical lysis of S. aureus prior to pathogen enrichment resulted in increased recovery of pathogen DNA. Quantification of pathogen and human DNA after selective lysis of blood cells and pathogen enrichment confirmed partial depletion of human DNA, leading to a net enrichment of pathogen DNA over human DNA. CONCLUSION: Concurrent antibiotic administration does not reduce the recovery of pathogen DNA during pathogen enrichment by selective lysis and centrifugation. Leads to a 10-fold human DNA depletion as compared to pathogen DNA. Moreover, we confirm that the recovery of pathogen DNA after pathogen enrichment is not negatively influenced by concurrent antibiotic administration. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12866-019-1559-7) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6683330 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-66833302019-08-09 Influence of antibiotic treatment on the detection of S. aureus in whole blood following pathogen enrichment Pilecky, Matthias Schildberger, Anita Knabl, Ludwig Orth-Höller, Dorothea Weber, Viktoria BMC Microbiol Methodology Article BACKGROUND: Early pathogen detection and identification are crucial for an effective and targeted antibiotic therapy in patients suffering from blood stream infection. Molecular diagnostic methods can accelerate pathogen identification as compared to blood culture, but frequently suffer from the inhibition of polymerase chain reation (PCR) by sample matrix components, such as host DNA, anticoagulants, or plasma proteins. To overcome this limitation, molecular diagnostic methods commonly rely on pathogen enrichment by selective lysis of blood cells and pelleting of intact pathogens prior to analysis. RESULTS: Here, we investigated the impact of antibiotic treatment on the recovery of pathogen DNA using an established pathogen enrichment protocol. Based on the hypothesis that induction of bacterial cell wall disintegration following antibiotic administration leads to incomplete pelleting of pathogen DNA, S. aureus was grown in human whole blood with or without addition of cell wall active (vancomycin, piperacillin) or non cell wall active (ciprofloxacin, clindamycin) antibiotics at clinically relevant concentrations. Pathogen detection remained unaffected by non cell wall active antibiotics or even increased in the presence of cell wall active antibiotics, indicating improved accessibility of pathogen DNA. Likewise, mechanical lysis of S. aureus prior to pathogen enrichment resulted in increased recovery of pathogen DNA. Quantification of pathogen and human DNA after selective lysis of blood cells and pathogen enrichment confirmed partial depletion of human DNA, leading to a net enrichment of pathogen DNA over human DNA. CONCLUSION: Concurrent antibiotic administration does not reduce the recovery of pathogen DNA during pathogen enrichment by selective lysis and centrifugation. Leads to a 10-fold human DNA depletion as compared to pathogen DNA. Moreover, we confirm that the recovery of pathogen DNA after pathogen enrichment is not negatively influenced by concurrent antibiotic administration. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12866-019-1559-7) contains supplementary material, which is available to authorized users. BioMed Central 2019-08-06 /pmc/articles/PMC6683330/ /pubmed/31387527 http://dx.doi.org/10.1186/s12866-019-1559-7 Text en © The Author(s). 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Methodology Article Pilecky, Matthias Schildberger, Anita Knabl, Ludwig Orth-Höller, Dorothea Weber, Viktoria Influence of antibiotic treatment on the detection of S. aureus in whole blood following pathogen enrichment |
title | Influence of antibiotic treatment on the detection of S. aureus in whole blood following pathogen enrichment |
title_full | Influence of antibiotic treatment on the detection of S. aureus in whole blood following pathogen enrichment |
title_fullStr | Influence of antibiotic treatment on the detection of S. aureus in whole blood following pathogen enrichment |
title_full_unstemmed | Influence of antibiotic treatment on the detection of S. aureus in whole blood following pathogen enrichment |
title_short | Influence of antibiotic treatment on the detection of S. aureus in whole blood following pathogen enrichment |
title_sort | influence of antibiotic treatment on the detection of s. aureus in whole blood following pathogen enrichment |
topic | Methodology Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6683330/ https://www.ncbi.nlm.nih.gov/pubmed/31387527 http://dx.doi.org/10.1186/s12866-019-1559-7 |
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