Please use this identifier to cite or link to this item: http://hdl.handle.net/1946/28785
Introduction: Elevated levels of cell free DNA (cf-DNA) in plasma is associated with various medical emergencies such as sepsis, stroke, trauma. In sepsis, levels of cf-DNA correlate with disease severity and hospital mortality25. The goal of this study was to examine if structural damage in cf-DNA is present in sepsis patients.
Materials and methods: Samples were collected from nine consenting patients (age 18+) admitted to the two ICU´s of Landspitali due to severe sepsis from December 2016 until May 2017. Samples were also collected from five healthy controls. The sample DNA was isolated using the Genomic Mini AX kit according to protocol. Northern Lights assay (NLA, two dimensional) was used to detect cf-DNA structural damage such as single/double strand breaks, bends, and inter/intra strand links. The addition of Fpg (formamidopyrimidine [fapy]-DNA glycosylase) which exaggerates damage was also examined for effect. The study was approved by the National Bioethics Committee and the Icelandic Data Authority.
Results: Elevated levels of cf-DNA were observed in sepsis patients which in turn lowered with treatment. Visual analysis of the NLA imaging results revealed a repeating pattern indicative of significant single-stranded breaks in nucleosomal DNA in sepsis patients that were not readily observable in controls. Fpg enzyme treatment decreased larger DNA molecules compared to control samples.
Conclusion: The increase in single-stranded breaks is possibly the result of lack of DNA repair associated with cell death. Larger DNA molecules that are isolated presumably derive from necrotic cells and possibly contained increased oxidative damage. The sample size in this study is too small to draw any concrete conclusions, and further research is warranted.
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