DNA Thermo-Protection Facilitates Whole-Genome Sequencing of Mycobacteria Direct from Clinical Samples
Sophie George, Yifei Xu, Gillian Rodger, Marcus Morgan, Nicholas D. Sanderson, Sarah Hoosdally, Samantha Thulborn, Esther Robinson, Priti Rathod, A Sarah Walker, Tim Peto, Derrick W. Crook, Kate E. Dingle
Abstract
Mycobacterium tuberculosis is the leading cause of death from bacterial infection. Improved rapid diagnosis and antimicrobial resistance determination, such as by whole-genome sequencing, are required. Our aim was to develop a simple, low-cost method of preparing DNA for sequencing direct from M. tuberculosis -positive clinical samples (without culture). Simultaneous sputum liquefaction, bacteria heat inactivation (99°C/30 min), and enrichment for mycobacteria DNA were achieved using an equal volume of thermo-protection buffer (4 M KCl, 0.
Topics & Concepts
GenomeBiologyDNA sequencingWhole genome sequencingDNAMycobacteriumMicrobiologyGeneticsComputational biologyBacteriaGeneMycobacterium research and diagnosisTuberculosis Research and EpidemiologyAntibiotic Resistance in Bacteria