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nuc DNA in Whole blood in Relation to Immune dysregulation, Sepsis, and Mortality in Staphylococcus aureus Bacteremia
Örebro University, School of Medical Sciences. Department of Infectious Diseases, Örebro University Hospital, Örebro, Sweden.
Örebro University, School of Medical Sciences. Örebro University Hospital. Department of Infectious Diseases.
Department of Infectious Diseases, Örebro University Hospital, Örebro, Sweden; School of Health and Medical Sciences, Örebro University, Örebro, Sweden.
Department of Laboratory Medicine, Örebro University Hospital, Örebro, Sweden.
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(English)Manuscript (preprint) (Other academic)
National Category
General Practice
Identifiers
URN: urn:nbn:se:oru:diva-68880OAI: oai:DiVA.org:oru-68880DiVA, id: diva2:1247537
Available from: 2018-09-12 Created: 2018-09-12 Last updated: 2018-09-12Bibliographically approved
In thesis
1. Quantitative detection of bacterial DNA in whole blood in bloodstream infection
Open this publication in new window or tab >>Quantitative detection of bacterial DNA in whole blood in bloodstream infection
2018 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

This thesis aims to increase the knowledge on how quantitative PCR can be used in the diagnostics of bloodstream infections, with an emphasis on quantitative elements.

In Papers I and II, we evaluated quantitative data from two commercial PCR tests for pathogen detection directly in blood, Magicplex Sepsis (I) and SeptiFast (II), from patients with suspected sepsis. We found that high quantification cycle (Cq) values, indicating low DNA loads, were associated with findings of pathogens with doubtful clinical relevance, whereas low Cq values, indicating high DNA loads, were correlated with sepsis and septic shock, as well as with positive blood culture results.

In Paper III, we aimed to study the bacterial DNA load during Staphylococcus aureus bacteremia, in relation to different clinical factors. For this purpose, we developed a droplet digital PCR (ddPCR) for precise DNA quantification, targeting S. aureus specifically. We found that a high initial S. aureus DNA load was associated with laboratory markers for immune dysregulation as well as with sepsis, endocarditis, and mortality.

In Paper IV, we aimed to develop a tool for repeated DNA quantification during bloodstream infection. For this purpose, we optimized a ddPCR, targeting the universal bacterial 16S rDNA, and performed a comparison with species-specific ddPCRs on spiked blood, and on clinical samples. The performance of the16S rDNA ddPCR was adequate, and we found that a high 16S rDNA load was associated with sepsis and mortality.

In conclusion, our results indicate that the pathogen DNA load in blood plays an important role in the clinical picture in BSI. In future research on molecular BSI diagnostics, studies on DNA loads and clearance should be included.

Place, publisher, year, edition, pages
Örebro: Örebro University, 2018. p. 94
Series
Örebro Studies in Medicine, ISSN 1652-4063 ; 183
Keywords
Bloodstream infection, bacteremia, sepsis, DNA load, quantitative PCR, droplet digital PCR, 16S rDNA
National Category
General Practice
Identifiers
urn:nbn:se:oru:diva-68452 (URN)978-91-7529-257-1 (ISBN)
Public defence
2018-10-05, Örebro universitet, Campus USÖ, hörsal C1, Södra Grev Rosengatan 32, Örebro, 13:00 (Swedish)
Opponent
Supervisors
Available from: 2018-08-14 Created: 2018-08-14 Last updated: 2018-09-13Bibliographically approved

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Ziegler, IngridCajander, Sara

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CiteExportLink to record
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