Sensitive Identification of Bacterial DNA in Clinical Specimens by Broad-Range 16S rRNA Gene Enrichment.

16S rRNA broad range enrichment hybridization capture metagenomics molecular diagnosis next-generation sequencing sequencing

Journal

Journal of clinical microbiology
ISSN: 1098-660X
Titre abrégé: J Clin Microbiol
Pays: United States
ID NLM: 7505564

Informations de publication

Date de publication:
18 11 2020
Historique:
received: 30 06 2020
accepted: 30 09 2020
pubmed: 9 10 2020
medline: 24 6 2021
entrez: 8 10 2020
Statut: epublish

Résumé

The broad-range detection and identification of bacterial DNA from clinical specimens are a foundational approach in the practice of molecular microbiology. However, there are circumstances under which conventional testing may yield false-negative or otherwise uninterpretable results, including the presence of multiple bacterial templates or degraded nucleic acids. Here, we describe an alternative, next-generation sequencing approach for the broad range detection of bacterial DNA using broad-range 16S rRNA gene hybrid capture ("16S Capture"). The method is able to deconvolute multiple bacterial species present in a specimen, is compatible with highly fragmented templates, and can be readily implemented when the overwhelming majority of nucleic acids in a specimen derive from the human host. We find that this approach is sensitive to detecting as few as 17

Identifiants

pubmed: 33028602
pii: JCM.01605-20
doi: 10.1128/JCM.01605-20
pmc: PMC7685877
pii:
doi:

Substances chimiques

DNA, Bacterial 0
RNA, Ribosomal, 16S 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Informations de copyright

Copyright © 2020 American Society for Microbiology.

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Auteurs

Sara Rassoulian Barrett (S)

Department of Laboratory Medicine and Pathology, University of Washington, Seattle, Washington, USA.

Noah G Hoffman (NG)

Department of Laboratory Medicine and Pathology, University of Washington, Seattle, Washington, USA.

Christopher Rosenthal (C)

Department of Laboratory Medicine and Pathology, University of Washington, Seattle, Washington, USA.

Andrew Bryan (A)

Department of Laboratory Medicine and Pathology, University of Washington, Seattle, Washington, USA.

Desiree A Marshall (DA)

Department of Laboratory Medicine and Pathology, University of Washington, Seattle, Washington, USA.

Joshua Lieberman (J)

Department of Laboratory Medicine and Pathology, University of Washington, Seattle, Washington, USA.

Alexander L Greninger (AL)

Department of Laboratory Medicine and Pathology, University of Washington, Seattle, Washington, USA.

Vikas Peddu (V)

Department of Laboratory Medicine and Pathology, University of Washington, Seattle, Washington, USA.

Brad T Cookson (BT)

Department of Laboratory Medicine and Pathology, University of Washington, Seattle, Washington, USA.
Department of Microbiology, University of Washington, Seattle, Washington, USA.

Stephen J Salipante (SJ)

Department of Laboratory Medicine and Pathology, University of Washington, Seattle, Washington, USA stevesal@uw.edu.

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Classifications MeSH