DNA extraction from primary liquid blood cultures for bloodstream infection diagnosis using whole genome sequencing.

ABSTRACT:

Purpose

Speed of bloodstream infection diagnosis is vital to reduce morbidity and mortality. Whole genome sequencing (WGS) performed directly from liquid blood culture could provide single-assay species and antibiotic susceptibility prediction; however, high inhibitor and human cell/DNA concentrations limit pathogen recovery. We develop a method for the preparation of bacterial DNA for WGS-based diagnostics direct from liquid blood culture.

Methodology

We evaluate three commercial DNA extraction kits: BiOstic Bacteraemia, Amplex Hyplex and MolYsis Plus. Differential centrifugation, filtration, selective lysis and solid-phase reversible immobilization bead clean-up are tested to improve human cells/DNA and inhibitor removal. Using WGS (Illumina/MinION), we assess human DNA re

SUBMITTER: Anson LW 

PROVIDER: S-EPMC5882078 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature