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A rapid high-throughput sequencing-based approach for the identification of unknown bacterial pathogens in whole blood

High-throughput DNA sequencing (HTS) of pathogens in whole blood samples is hampered by the high host/pathogen nucleic acids ratio. We describe a novel and rapid bacterial enrichment procedure whose implementation is exemplified in simulated bacteremic human blood samples. The procedure involves dep...

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Detalles Bibliográficos
Autores principales: Israeli, Ofir, Makdasi, Efi, Cohen-Gihon, Inbar, Zvi, Anat, Lazar, Shirley, Shifman, Ohad, Levy, Haim, Gur, David, Laskar, Orly, Beth-Din, Adi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Future Science Ltd 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7351085/
https://www.ncbi.nlm.nih.gov/pubmed/32670604
http://dx.doi.org/10.2144/fsoa-2020-0013
Descripción
Sumario:High-throughput DNA sequencing (HTS) of pathogens in whole blood samples is hampered by the high host/pathogen nucleic acids ratio. We describe a novel and rapid bacterial enrichment procedure whose implementation is exemplified in simulated bacteremic human blood samples. The procedure involves depletion of the host DNA, rapid HTS and bioinformatic analyses. Following this procedure, Y. pestis, F. tularensis and B. anthracis spiked-in samples displayed an improved host/pathogen DNA ratio of 2.5–5.9 orders of magnitude, in samples with bacteria spiked-in at 10(3)–10(5) CFU/ml. The procedure described in this study enables rapid and detailed metagenomic profiling of pathogens within 8–9 h, circumventing the challenges imposed by the high background present in the bacteremic blood and by the unknown nature of the sample.