Microplate alamar blue assay versus BACTEC 460 system for high-throughput screening of compounds against Mycobacterium tuberculosis and Mycobacterium avium
Open accessAbstract
In response to the need for rapid, inexpensive, high-throughput assays for antimycobacterial drug screening, a microplate-based assay which uses Alamar blue reagent for determination of growth was evaluated. MICs of 30 antimicrobial agents against Mycobacterium tuberculosis H37Rv, M. tuberculosis H37Ra, and Mycobacterium avium were determined in the microplate Alamar blue assay (MABA) with both visual and fluorometric readings and compared to MICs determined in the BACTEC 460 system. For all three mycobacterial strains, there was < or = 1 dilution difference between MABA and BACTEC median MICs in four replicate experiments for 25 to 27 of the 30 antimicrobics. Significant differences between MABA and BACTEC MICs were observed with 0, 2, and 5 of 30 antimicrobial agents against H37Rv, H37Ra, and M. avium, respectively. Overall, MICs determined either visually or fluorometrically in MABA were highly correlated with those determined in the BACTEC 460 system, and visual MABA and fluorometric MABA MICs were highly correlated. MICs of rifampin, rifabutin, minocycline, and clarithromycin were consistently lower for H37Ra compared to H37Rv in all assays but were similar for most other drugs. M. tuberculosis H37Ra may be a suitable surrogate for the more virulent H37Rv strain in primary screening of compounds for antituberculosis activity. MABA is sensitive, rapid, inexpensive, and nonradiometric and offers the potential for screening, with or without analytical instrumentation, large numbers of antimicrobial compounds against slow-growing mycobacteria.
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MDR TB is most prevalent in some countries of Eastern Europe as well as China and India.
“MDR TB, defined by resistance to at least isoniazid and rifampin, is ubiquitous (12) and is most prevalent in some countries of Eastern Europe as well as China and India (14, 47).”
From Resazurin Microtiter Assay Plate Testing of <i>Mycobacterium tuberculosis</i> Susceptibilities to Second-Line Drugs: Rapid, Simple, and Inexpensive Method (2003) · cites this paper as [14]
The paper’s introduction mentions the need for drugs against multidrug-resistant TB, but gives no country-level prevalence figures. The Discussion concerns drug-susceptibility assays and bacterial strains, not geographic prevalence; searches for Eastern Europe, China, India, and geographic distribution found no supporting passage in our copy. (LLM, conf=0.76)
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- Authors
- Collins, L., & Franzblau, S. G.
- Affiliations
- Pharmacology Research Department, Gillis W. Long Hansen's Disease Center, Baton Rouge, Louisiana 70894, USA.
- ORCID
- Franzblau, S G
- Venue
- Antimicrobial Agents and Chemotherapy
- Publisher
- American Society for Microbiology
- Published
- 1997-05
- Volume
- 41
- Issue
- 5
- Pages
- 1004-1009
- ISSN
- 0066-4804 (ISSN-L)
- 1098-6596
- PMC
- PMC163841
- OpenAlex
- W2126576359
- Cited by
- 1,599
- References
- 0
- Citations by year
- 2026: 18 · 2025: 31 · 2024: 35 · 2023: 44 · 2022: 46 · 2021: 48 · 2020: 53 · 2019: 49 · 2018: 65 · 2017: 63 · 2016: 69 · 2015: 73 · 2014: 74 · 2013: 84 · 2012: 98
- FWCI
- 13.41
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Antimicrobial Agents and Chemotherapy (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institute of Allergy and Infectious Diseases (Y02-AI-50101-00)
- Topics
- Health Sciences › Medicine › Infectious Diseases › Tuberculosis Research and Epidemiology
- Health Sciences › Medicine › Epidemiology › Mycobacterium research and diagnosis
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Clinical Biochemistry › Bacterial Identification and Susceptibility Testing
- Keywords
- Alamar Blue assay, BACTEC 460, Mycobacterium tuberculosis, Mycobacterium avium, minimum inhibitory concentration, high-throughput screening, Mycobacterium tuberculosis H37Rv, H37Ra, rifampicin, rifabutin, minocycline, clarithromycin
- MeSH terms
- Anti-Bacterial Agents, Drug Evaluation, Preclinical, Coloring Agents, Fluorometry, Microbial Sensitivity Tests, Mycobacterium avium, Mycobacterium tuberculosis, Oxazines, Radiometry, Xanthenes
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