Removing the bottleneck in whole genome sequencing of Mycobacterium tuberculosis for rapid drug resistance analysis: a call to action

R McNerney, TG Clark, S Campino, C Rodrigues, D Dolinger, L Smith, AM Cabibbe, K Dheda, Marco Schito

Research output: Contribution to journalArticle

Abstract

Whole genome sequencing (WGS) can provide a comprehensive analysis of Mycobacterium tuberculosis mutations that cause resistance to anti-tuberculosis drugs. With the deployment of bench-top sequencers and rapid analytical software, WGS is poised to become a useful tool to guide treatment. However, direct sequencing from clinical specimens to provide a full drug resistance profile remains a serious challenge. This article reviews current practices for extracting M. tuberculosis DNA and possible solutions for sampling sputum. Techniques under consideration include enzymatic digestion, physical disruption, chemical degradation, detergent solubilization, solvent extraction, ligand-coated magnetic beads, silica columns, and oligonucleotide pull-down baits. Selective amplification of genomic bacterial DNA in sputum prior to WGS may provide a solution, and differential lysis to reduce the levels of contaminating human DNA is also being explored. To remove this bottleneck and accelerate access to WGS for patients with suspected drug-resistant tuberculosis, it is suggested that a coordinated and collaborative approach be taken to more rapidly optimize, compare, and validate methodologies for sequencing from patient samples. © 2016 The Author(s)
Original languageEnglish
Pages (from-to)130-135
Number of pages6
JournalInternational Journal of Infectious Diseases
Volume56
Issue number16
DOIs
Publication statusPublished - 2017

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    McNerney, R., Clark, TG., Campino, S., Rodrigues, C., Dolinger, D., Smith, L., Cabibbe, AM., Dheda, K., & Schito, M. (2017). Removing the bottleneck in whole genome sequencing of Mycobacterium tuberculosis for rapid drug resistance analysis: a call to action. International Journal of Infectious Diseases, 56(16), 130-135. https://doi.org/10.1016/j.ijid.2016.11.422