DNA Thermo-Protection Facilitates Whole-Genome Sequencing of Mycobacteria Direct from Clinical Samples-Reference-Cited by-同舟云学术

DNA Thermo-Protection Facilitates Whole-Genome Sequencing of Mycobacteria Direct from Clinical Samples

Published:2020-09-22 Issue:10 Volume:58 Page:
ISSN:0095-1137
Container-title:Journal of Clinical Microbiology
language:en
Short-container-title:J Clin Microbiol

Author:

George Sophie¹²,Xu Yifei¹²,Rodger Gillian¹²,Morgan Marcus³,Sanderson Nicholas D.¹²,Hoosdally Sarah J.¹²,Thulborn Samantha²⁴,Robinson Esther⁵,Rathod Priti⁵,Walker A. Sarah¹²,Peto Timothy E. A.¹²,Crook Derrick W.¹²,Dingle Kate E.¹²^ORCID

Affiliation:

1. Nuffield Department of Clinical Medicine, John Radcliffe Hospital, Oxford University, Oxford, United Kingdom

2. National Institute for Health Research (NIHR) Oxford Biomedical Research Centre, John Radcliffe Hospital, Oxford, United Kingdom

3. Microbiology Department, Oxford University Hospitals NHS Trust, Oxford, United Kingdom

4. Respiratory Medicine Unit, Nuffield Department of Medicine, John Radcliffe Hospital, University of Oxford, Oxford, United Kingdom

5. PHE National Mycobacteria Reference Service—North and Central, Birmingham Public Health Laboratory, Birmingham, United Kingdom

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.

Funder

NIHR | NIHR Oxford Biomedical Research Centre

Publisher

American Society for Microbiology

Subject

Microbiology (medical)

Link

https://journals.asm.org/doi/pdf/10.1128/JCM.00670-20

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