1. Microbiology and Infectious Disease
Download icon

ODELAM Rapid sequence-independent detection of drug resistance in isolates of Mycobacterium tuberculosis

  1. Thurston Herricks
  2. Magdalena Donczew  Is a corresponding author
  3. Fred D Mast
  4. Tige Rustad
  5. Robert Morrison
  6. Timothy R Sterling
  7. David R Sherman  Is a corresponding author
  8. John D Aitchison  Is a corresponding author
  1. Seattle Children's Research Institute, United States
  2. University of Washington, United States
  3. Vanderbilt University, United States
Tools and Resources
  • Cited 4
  • Views 874
  • Annotations
Cite this article as: eLife 2020;9:e56613 doi: 10.7554/eLife.56613

Abstract

Antimicrobial-resistant Mycobacterium tuberculosis (Mtb) causes over 200,000 deaths each year. Current assays of antimicrobial resistance need knowledge of mutations that confer drug resistance, or long periods of culture time to test growth under drug pressure. We present ODELAM (One-cell Doubling Evaluation of Living Arrays of Mycobacterium), a time-lapse microscopy-based method that observes individual cells growing into microcolonies. ODELAM enables rapid quantitative measures of growth kinetics in as little as 30 hours under a wide variety of environmental conditions. We demonstrate ODELAM's utility by identifying ofloxacin resistance in cultured clinical isolates of Mtb and benchmark its performance with standard minimum inhibitory concentration (MIC) assays. In Mtb isolate, ODELAM identified ofloxacin heteroresistance and identifies the presence of drug resistant colony forming units (CFUs) at 1 per 1000 CFUs in as little as 48 hours. ODELAM is a powerful new tool that can rapidly evaluate Mtb drug resistance in a laboratory setting.

Data availability

MATLAB data *.mat files and MATLAB *.m files utilized for generating figures in this submission are posted at Dryad. Additional source code has been made available at https://github.com/AitchisonLab/

The following data sets were generated

Article and author information

Author details

  1. Thurston Herricks

    Center for Global Infectious Disease Research, Seattle Children's Research Institute, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0247-7967
  2. Magdalena Donczew

    Department of Microbiology, University of Washington, Seattle, United States
    For correspondence
    mdonczew@uw.edu
    Competing interests
    The authors declare that no competing interests exist.
  3. Fred D Mast

    Center for Global Infectious Disease Research, Seattle Children's Research Institute, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Tige Rustad

    Center for Global Infectious Disease Research, Seattle Children's Research Institute, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Robert Morrison

    Center for Global Infectious Disease Research, Seattle Children's Research Institute, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Timothy R Sterling

    Division of Infectious Disease, Department of Medicine, Vanderbilt University, Nashville, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. David R Sherman

    Department of Microbiology, University of Washington, Seattle, United States
    For correspondence
    dsherman@uw.edu
    Competing interests
    The authors declare that no competing interests exist.
  8. John D Aitchison

    Center for Global Infectious Disease Research, Seattle Children's Research Institute, Seattle, United States
    For correspondence
    John.Aitchison@seattlechildrens.org
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9153-6497

Funding

National Institutes of Health (U19 AI135976)

  • David R Sherman
  • John D Aitchison

National Institutes of Health (U19 AI111276)

  • John D Aitchison

National Institutes of Health (R01 AI141953)

  • John D Aitchison

National Institutes of Health (P41 GM109824)

  • John D Aitchison

National Institutes of Health (R01 AI063200)

  • Timothy R Sterling

National Institutes of Health (R56 AI118361)

  • Timothy R Sterling

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Miles P Davenport, University of New South Wales, Australia

Publication history

  1. Received: March 3, 2020
  2. Accepted: May 12, 2020
  3. Accepted Manuscript published: May 13, 2020 (version 1)
  4. Version of Record published: June 1, 2020 (version 2)

Copyright

© 2020, Herricks et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 874
    Page views
  • 148
    Downloads
  • 4
    Citations

Article citation count generated by polling the highest count across the following sources: PubMed Central, Crossref, Scopus.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Download citations (links to download the citations from this article in formats compatible with various reference manager tools)

Open citations (links to open the citations from this article in various online reference manager services)

Further reading

    1. Biochemistry and Chemical Biology
    2. Microbiology and Infectious Disease
    Giulia Bandini et al.
    Research Article Updated

    Fucose is a common component of eukaryotic cell-surface glycoconjugates, generally added by Golgi-resident fucosyltransferases. Whereas fucosylated glycoconjugates are rare in kinetoplastids, the biosynthesis of the nucleotide sugar GDP-Fuc has been shown to be essential in Trypanosoma brucei. Here we show that the single identifiable T. brucei fucosyltransferase (TbFUT1) is a GDP-Fuc: β-D-galactose α-1,2-fucosyltransferase with an apparent preference for a Galβ1,3GlcNAcβ1-O-R acceptor motif. Conditional null mutants of TbFUT1 demonstrated that it is essential for both the mammalian-infective bloodstream form and the insect vector-dwelling procyclic form. Unexpectedly, TbFUT1 was localized in the mitochondrion of T. brucei and found to be required for mitochondrial function in bloodstream form trypanosomes. Finally, the TbFUT1 gene was able to complement a Leishmania major mutant lacking the homologous fucosyltransferase gene (Guo et al., 2021). Together these results suggest that kinetoplastids possess an unusual, conserved and essential mitochondrial fucosyltransferase activity that may have therapeutic potential across trypanosomatids.

    1. Microbiology and Infectious Disease
    Michael J Sheedlo et al.
    Research Article

    Legionella pneumophila is an opportunistic pathogen that causes the potentially fatal pneumonia known as Legionnaires' Disease. The pathology associated with infection depends on bacterial delivery of effector proteins into the host via the membrane spanning Dot/Icm type IV secretion system (T4SS). We have determined sub-3.0 Å resolution maps of the Dot/Icm T4SS core complex by single particle cryo-EM. The high-resolution structural analysis has allowed us to identify proteins encoded outside the Dot/Icm genetic locus that contribute to the core T4SS structure. We can also now define two distinct areas of symmetry mismatch, one that connects the C18 periplasmic ring (PR) and the C13 outer membrane cap (OMC) and one that connects the C13 OMC with a 16-fold symmetric dome. Unexpectedly the connection between the PR and OMC is DotH, with five copies sandwiched between the OMC and PR to accommodate the symmetry mismatch. Finally, we observe multiple conformations in the reconstructions that indicate flexibility within the structure.