A Multi-stress Model for High Throughput Screening Against Non-replicating Mycobacterium tuberculosis

  • Ben GoldEmail author
  • Thulasi Warrier
  • Carl Nathan
Part of the Methods in Molecular Biology book series (MIMB, volume 1285)


Models of non-replication help us understand the biology of persistent Mycobacterium tuberculosis. High throughput screening (HTS) against non-replicating M. tuberculosis may lead to identification of tool compounds that affect pathways on which bacterial survival depends in such states, and to development of drugs that can overcome phenotypic tolerance to conventional antimycobacterial agents, which are mostly active against replicating M. tuberculosis. We describe a multi-stress model of non-replication that mimics some of the microenvironmental conditions that M. tuberculosis faces in the host as adapted for HTS. The model includes acidic pH, mild hypoxia, a flux of nitric oxide and other reactive nitrogen intermediates arising from nitrite at low pH, and low concentrations of a fatty acid (butyrate) as a carbon source.

Key words

Mycobacterium tuberculosis Non-replication Hypoxia Acidic pH Fatty acids Nitric oxide Reactive nitrogen species Phenotypic tolerance Antibiotics High-throughput screening 



We are very grateful for the numerous contributions towards the development of the non-replicating assay and critical reading of this chapter: Julia Roberts, Yan Ling, Selin Somersan-Karakaya, Landys Lopez-Quezada, David Little, Maneesh Pingle, Kristin Burns-Huang, and Alfonso Mendoza-Losana, Maria Martinez-Hoyos, and Manuel Marin-Amieva at GSK in Tres Cantos, Spain. This work was supported by the TB Drug Accelerator Program of the Bill and Melinda Gates Foundation and by the Abby and Howard P. Milstein Program in Translational Medicine. The Department of Microbiology and Immunology is supported by the William Randolph Hearst Foundation.


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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of Microbiology and ImmunologyWeill Cornell Medical CollegeNew YorkUSA
  2. 2.Department of Microbiology and ImmunologyWeill Cornell Medical CollegeNew YorkUSA
  3. 3.Department of Microbiology and ImmunologyWeill Cornell Medical CollegeNew YorkUSA

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