Quantification of bacterial DNA in blood using droplet digital PCR: a pilot study.

Autor: Tedim AP; Group for Biomedical Research in Sepsis (BioSepsis), Instituto de Investigación Biomédica de Salamanca, (IBSAL), Salamanca, Spain; Hospital Universitario Río Hortega, Valladolid, Spain; Centro de Investigación Biomédica en Red en Enfermedades Respiratorias (CiberES), CB22/06/00035, Instituto de Salud Carlos III, Madrid, Spain. Electronic address: anspedrosa@gmail.com., Merino I; Microbiology Department, Hospital Universitario Río Hortega, Valladolid, Spain., Ortega A; Group for Biomedical Research in Sepsis (BioSepsis), Instituto de Investigación Biomédica de Salamanca, (IBSAL), Salamanca, Spain; Hospital Universitario Río Hortega, Valladolid, Spain; Centro de Investigación Biomédica en Red en Enfermedades Respiratorias (CiberES), CB22/06/00035, Instituto de Salud Carlos III, Madrid, Spain., Domínguez-Gil M; Microbiology Department, Hospital Universitario Río Hortega, Valladolid, Spain., Eiros JM; Microbiology Department, Hospital Universitario Río Hortega, Valladolid, Spain., Bermejo-Martín JF; Group for Biomedical Research in Sepsis (BioSepsis), Instituto de Investigación Biomédica de Salamanca, (IBSAL), Salamanca, Spain; Hospital Universitario Río Hortega, Valladolid, Spain; Centro de Investigación Biomédica en Red en Enfermedades Respiratorias (CiberES), CB22/06/00035, Instituto de Salud Carlos III, Madrid, Spain.
Jazyk: angličtina
Zdroj: Diagnostic microbiology and infectious disease [Diagn Microbiol Infect Dis] 2024 Jan; Vol. 108 (1), pp. 116075. Date of Electronic Publication: 2023 Sep 04.
DOI: 10.1016/j.diagmicrobio.2023.116075
Abstrakt: We used droplet digital PCR (ddPCR) assays to detect/quantify DNA from Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, and Enterococcus spp. in blood samples. Bacterial DNA from clinical strains (4 < n < 12) was extracted, quantified and diluted (10-0.0001 ng/µL) and ddPCR assays were performed in triplicate. These ddPCR assays showed low replication variability, low detection limit (1-0.1 pg/µL), and genus/species specificity. ddPCR assays were also used to quantify bacterial DNA obtained from spiked blood (1 × 10 4 -1 CFU/mL) of each bacterial genus/species. Comparison between ddPCR assays and bacterial culture was performed by Pearson correlation. There was an almost perfect correlation (r ≥ 0.997, P ≤ 0.001) between the number of CFU/mL from bacterial culture and the number of gene copies/mL detected by ddPCR. The time from sample preparation to results was determined to be 3.5 to 4 hours. The results demonstrated the quantification capacity and specificity of the ddPCR assays to detect/quantify 4 of the most important bloodstream infection (BSI) bacterial pathogens directly from blood. SIGNIFICANCE AND IMPACT: This pilot study results support the potential of ddPCR for the diagnosis and/or severity stratification of BSI. Applied to patients' blood samples it can improve diagnosis and diminish sample-to-results time, improving patient care.
Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Ana P. Tedim reports financial support was provided by Carlos III Health Institute. Ana P. Tedim reports financial support was provided by European Society of Clinical Microbiology and Infectious Diseases. Jesus Francisco Bermejo Martin reports financial support was provided by Carlos III Health Institute.
(Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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