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. 2017 Feb;36(2):329-336.
doi: 10.1007/s10096-016-2805-7. Epub 2016 Oct 22.

Comprehensive detection and identification of bacterial DNA in the blood of patients with sepsis and healthy volunteers using next-generation sequencing method - the observation of DNAemia

T Gosiewski  1 , A H Ludwig-Galezowska  2 , K Huminska  3   4 , A Sroka-Oleksiak  1 , P Radkowski  2 , D Salamon  1 , J Wojciechowicz  3 , M Kus-Slowinska  3 , M Bulanda  1 , P P Wolkow  5
Affiliations

Comprehensive detection and identification of bacterial DNA in the blood of patients with sepsis and healthy volunteers using next-generation sequencing method - the observation of DNAemia

T Gosiewski et al. Eur J Clin Microbiol Infect Dis. 2017 Feb.

Abstract

Blood is considered to be a sterile microenvironment, in which bacteria appear only periodically. Previously used methods allowed only for the detection of either viable bacteria with low sensitivity or selected species of bacteria. The Next-Generation Sequencing method (NGS) enables the identification of all bacteria in the sample with their taxonomic classification. We used NGS for the analysis of blood samples from healthy volunteers (n = 23) and patients with sepsis (n = 62) to check whether any bacterial DNA exists in the blood of healthy people and to identify bacterial taxonomic profile in the blood of septic patients. The presence of bacterial DNA was found both in septic and healthy subjects; however, bacterial diversity was significantly different (P = 0.002) between the studied groups. Among healthy volunteers, a significant predominance of anaerobic bacteria (76.2 %), of which most were bacteria of the order Bifidobacteriales (73.0 %), was observed. In sepsis, the majority of detected taxa belonged to aerobic or microaerophilic microorganisms (75.1 %). The most striking difference was seen in the case of Actinobacteria phyla, the abundance of which was decreased in sepsis (P < 0.001) and Proteobacteria phyla which was decreased in the healthy volunteers (P < 0.001). Our research shows that bacterial DNA can be detected in the blood of healthy people and that its taxonomic composition is different from the one seen in septic patients. Detection of bacterial DNA in the blood of healthy people may suggest that bacteria continuously translocate into the blood, but not always cause sepsis; this observation can be called DNAemia.

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Conflict of interest statement

Compliance with ethical standards Conflicts of interest The authors declare no conflicts of interest. Ethical approval All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. The research was granted approval by the local Bioethics Committee of the Jagiellonian University (KBET/94/B/2009). Written informed consent was obtained from participants before their enrollment in the study.

Figures

Fig. 1
Fig. 1
Rarefaction curves for patients with sepsis (orange curve) versus healthy patients (red curve) and NTC samples (blue curve); p = 0.002
Fig. 2
Fig. 2
Weighted UniFrac PCoA plot derived from NGS sequencing of blood samples taken from healthy volunteers (n = 23, red), NTC samples (blue) and patients with sepsis (n = 62, yellow)
Fig. 3
Fig. 3
Phyla abundances in the studied groups
Fig. 4
Fig. 4
Effect of patients status on taxonomy composition of order abundances
Fig. 5
Fig. 5
Representative results of nested amplification of the 16S libraries and the negative control (NTC). Amplicon 550 bp
See this image and copyright information in PMC

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