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Cited by 115Tuberculosis Research and Epidemiologybioluminescence and chemiluminescence researchMycobacterium research and diagnosis

Rapid measurement of antituberculosis drug activity in vitro and in macrophages using bioluminescence

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Andreu, N., Fletcher, T., Krishnan, N., Wiles, S., Robertson, B. D.·2012·Journal of Antimicrobial Chemotherapy
DOI 10.1093/jac/dkr472PMC PMC3254196
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Abstract

OBJECTIVES: Tuberculosis drug development is hampered by the slow growth of Mycobacterium tuberculosis. Bioluminescence, light produced by an enzymatic reaction, constitutes a rapid and highly sensitive measurement of cell metabolic function that can be used as an indirect marker of cell viability in drug screening assays. The aim of this work was to validate and standardize the use of luminescent M. tuberculosis strains to test the activity of antibacterial drugs in vitro and inside macrophages in a 96-well format. METHODS: We have used strains that express the bacterial lux operon and therefore do not require exogenous substrate to produce light, as well as strains expressing the firefly luciferase that need luciferin substrate. Results were compared with those obtained using the resazurin reduction assay and cfu plating. RESULTS: Using bioluminescence we were able to reduce the time required to measure the MIC and bactericidal concentrations of antimicrobials to just 3 and 6 days, respectively. Furthermore, antibacterial activity against intracellular mycobacteria was detected within 2 days post-infection. Results were comparable to those obtained by conventional methods. CONCLUSIONS: We have developed a simple and rapid method for screening antimycobacterial drugs in culture and in macrophages. The use of autoluminescent bacteria also facilitates the determination of growth and inhibition kinetics. The method is cost-effective, can easily be adapted to a larger scale and is amenable to automation. Current efforts are directed towards applying this technology to drug screening in vivo.

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  • The ATP bioluminescence technique can provide results in 3-5 days for antimycobacterial tests in comparison with the conventional dilution technique, which requires 3-4 weeks of incubation.

    “Indeed it has been demonstrated that this technique can provide results in 3–5 days for antimycobacterial tests [90], [91] in comparison with the conventional dilution technique, which requires 3–4 weeks of incubation [90,91].”

    From Methods for in vitro evaluating antimicrobial activity: A review (2016) · cites this paper as [90]

    The Abstract Results report that bioluminescence yielded MIC results in 3 days, supporting the claim of faster antimycobacterial testing. But the Introduction assigns the 3–5-day range to an optical-density microdilution assay and the 3–4-week period to colony formation on solid medium, not to the bioluminescence and conventional dilution techniques, respectively. The paper does not identify its method as ATP bioluminescence. (LLM, conf=0.88)

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  • Bioluminescence assay has been used by several authors for antibacterial testing, antimycobacterial testing, antifungal against yeast and molds.

    “Moreover, It has been used by several authors for antibacterial testing [89], antimycobacterial testing [90], [91], antifungal against yeast [92] and molds [93].”

    From Methods for in vitro evaluating antimicrobial activity: A review (2016) · cites this paper as [90]

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Research works printed but not linked: 32

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Authors
Andreu, N., Fletcher, T., Krishnan, N., Wiles, S., & Robertson, B. D.
Affiliations
  • Imperial College London (GB)
  • University of Auckland (NZ)
Corresponding author
Wiles, S.
ORCID
Venue
Journal of Antimicrobial Chemotherapy
Publisher
Oxford University Press (OUP)
Published
2011-11-17
Volume
67
Issue
2
Pages
404-414
ISSN
  • 0305-7453 (ISSN-L)
  • 1460-2091
DOI
10.1093/jac/dkr472
PMC
PMC3254196
OpenAlex
W2137161479
Cited by
115
References
0
Citations by year
2026: 4 · 2025: 3 · 2024: 8 · 2023: 2 · 2022: 4 · 2021: 17 · 2020: 8 · 2019: 12 · 2018: 6 · 2017: 7 · 2016: 23 · 2015: 4 · 2014: 8 · 2013: 5 · 2012: 4
FWCI
3.68
Type
journal article
Language
en
Open access
Licence
cc-by-nc
Locations
  • Journal of Antimicrobial Chemotherapy (publishedVersion, cc-by-nc)
  • PubMed Central (publishedVersion, cc-by-nc)
  • PubMed (publishedVersion)
  • LSHTM Research Online (London School of Hygiene and Tropical Medicine) (acceptedVersion)
  • HighWire Press Open Archive (submittedVersion)
Funders
  • Bill and Melinda Gates Foundation
  • Biotechnology and Biological Sciences Research Council (BB/E52708X/1)
  • Health Research Council of New Zealand (09/099)
Topics
  • Health Sciences › Medicine › Infectious Diseases › Tuberculosis Research and Epidemiology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › bioluminescence and chemiluminescence research
  • Health Sciences › Medicine › Epidemiology › Mycobacterium research and diagnosis
Keywords
Mycobacterium tuberculosis, bioluminescence, antimycobacterial drugs, macrophages, intracellular mycobacteria, lux operon, firefly luciferase, minimum inhibitory concentration, bactericidal concentration, 96-well assay, resazurin reduction assay
MeSH terms
Antitubercular Agents, Costs and Cost Analysis, Humans, Luciferases, Luminescent Measurements, Macrophages, Microbial Sensitivity Tests, Mycobacterium tuberculosis, Sensitivity and Specificity, Time Factors, Genes, Reporter

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