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Assessing nanopore sequencing for clinical diagnostics: A comparison of NGS methods for Mycobacterium tuberculosis

Publication

Date: 14th October 2020 | Source: Journal of Clinical Microbiology

Authors: Carol Smith, Tanya A. Halse, Joseph Shea, Herns Modestil, Randal C. Fowler, Kimberlee A. Musser, Vincent Escuyer, Pascal Lapierre.

Next-generation sequencing technologies are being rapidly adopted as a tool of choice for diagnostic and outbreak investigation in public health laboratories. However, costs of operation and the need for specialized staff remain major hurdles for laboratories with limited resources for implementing these technologies.

This project aimed to assess the feasibility of using Oxford Nanopore MinION whole-genome sequencing data of Mycobacterium tuberculosis isolates for species identification, in silico spoligotyping, detection of mutations associated with antimicrobial resistance (AMR) to accurately predict drug susceptibility profiles, and phylogenetic analysis to detect transmission between cases.

The results were compared prospectively in real-time, to those obtained with our current clinically validated Illumina MiSeq sequencing assay for M. tuberculosis and phenotypic drug susceptibility testing results when available. Our assessment of 431 sequenced samples over a 32-week period demonstrates that, when using the proper quality controls and thresholds, the MinION can achieve levels of genotyping analysis and phenotypic resistance predictions comparable to those of the Illumina MiSeq, at a very competitive cost per sample.

Our results indicate that nanopore sequencing can be a suitable alternative to, or complement, currently used sequencing platforms in a clinical setting and has the potential to be widely adopted in public health laboratories in the near future.

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