Rapid single-colony whole-genome sequencing of bacterial pathogens.

ABSTRACT:

Objectives

As a result of the introduction of rapid benchtop sequencers, the time required to subculture a bacterial pathogen to extract sufficient DNA for library preparation can now exceed the time to sequence said DNA. We have eliminated this rate-limiting step by developing a protocol to generate DNA libraries for whole-genome sequencing directly from single bacterial colonies grown on primary culture plates.

Methods

We developed our protocol using single colonies of 17 bacterial pathogens responsible for severe human infection that were grown using standard diagnostic media and incubation conditions. We then applied this method to four clinical scenarios that currently require time-consuming reference laboratory tests: full identification and genotyping of salmonellae;

SUBMITTER: Koser CU 

PROVIDER: S-EPMC3977605 | biostudies-literature | 2014 May

REPOSITORIES: biostudies-literature