16S rDNA droplet digital PCR for monitoring bacterial DNAemia in bloodstream infections


Autoři: Ingrid Ziegler aff001;  Sofia Lindström aff003;  Magdalena Källgren aff004;  Kristoffer Strålin aff002;  Paula Mölling aff002
Působiště autorů: Department of Infectious Diseases, Örebro University Hospital, Örebro, Sweden aff001;  School of Health and Medical Sciences, Örebro University, Örebro, Sweden aff002;  Department of Laboratory Medicine, Västerås Hospital, Västerås, Sweden aff003;  Department of Laboratory Medicine, Karlskoga Hospital, Karlskoga, Sweden aff004;  Department of Infectious Diseases, Karolinska University Hospital, Stockholm, Sweden aff005;  Department of Medicine Huddinge, Karolinska Institutet, Stockholm, Sweden aff006;  Department of Laboratory Medicine, Örebro University Hospital, Örebro, Sweden aff007
Vyšlo v časopise: PLoS ONE 14(11)
Kategorie: Research Article
doi: 10.1371/journal.pone.0224656

Souhrn

Repeated quantitative measurement of bacterial DNA on whole blood has been shown to be a promising method for monitoring bloodstream infection (BSI) with selected bacterial species. To enable broad use of this method, we developed a quantitative droplet digital PCR (ddPCR) method for 16S rDNA. It was validated with species-specific ddPCRs for Staphylococcus aureus (nuc), Streptococcus pneumoniae (lytA), and Escherichia coli (uidA) on spiked whole blood samples and on repeated whole blood samples (days 0, 1–2, 3–4, 6–8, and 13–15) from 83 patients with BSI with these pathogens. In these patients, 16S rDNA and species-specific DNA were detected in 60% and 61%, respectively, at least at one time-point. The highest positivity rates were seen in S. aureus BSI, where 92% of the patients were 16S rDNA-positive and 85% nuc-positive. Quantitative 16S rDNA and species-specific DNA showed strong correlations in spiked samples (r = 0.98; p < 0.0001) and clinical samples (r = 0.84; p < 0.0001). Positivity for 16S rDNA was rapidly cleared in patients with S. pneumoniae and E. coli BSI, but more slowly and sometimes persisted, in those with S. aureus BSI. The initial 16S rDNA load was higher in BSI patients with sepsis (Sepsis-3 definition) than without sepsis (median 2.38 vs. 0 lg10 copies/mL; p = 0.031) and in non-survivors than in survivors (median 2.83 vs. 0 lg10 copies/mL; p = 0.006). 16S rDNA ddPCR appears to be a promising method for bacterial DNA monitoring during BSI. The clinical value of such monitoring should be further studied.

Klíčová slova:

Bacterial pathogens – Blood – Bloodstream infections – Escherichia coli – Pneumococcus – Polymerase chain reaction – Sepsis – Staphylococcus aureus


Zdroje

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