Rapid, Low-Technology MIC Determination with Clinical Mycobacterium tuberculosis Isolates by Using the Microplate Alamar Blue Assay
Open accessAbstract
A colorimetric, microplate-based Alamar Blue assay (MABA) method was used to determine the MICs of isoniazid (INH), rifampin, streptomycin (SM), and ethambutol (EMB) for 34 Peruvian Mycobacterium tuberculosis isolates (including both pansensitive and multidrug-resistant strains) and the H 37 Rv strain by using bacterial suspensions prepared directly from solid media. Results for all isolates were available within 8 days. Discordant results were observed on initial tests for 3 of 16 INH-susceptible isolates, 5 of 31 EMB-susceptible isolates, and 2 of 4 SM-resistant isolates (by the BACTEC 460 system). The overall agreements between the MICs obtained by MABA and the results obtained with the BACTEC 460 system were 87.9% for initial results and 93.6% after retesting 12 of 17 samples with discrepant results. Interpretation of MABA endpoints improved with technical experience. The MABA is a simple, rapid, low-cost, appropriate technology which does not require expensive instrumentation and which makes use of a nontoxic, temperature-stable reagent.
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Preliminary studies of DST of MDR TB isolates using MTT, Alamar blue, or resazurin showed a good correlation with the conventional proportion method.
“Preliminary studies of DST of MDR TB isolates using MTT, Alamar blue, or resazurin showed a good correlation with the conventional PM (1, 13, 19, 20, 31, 37, 49).”
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 [20]
The paper’s introduction reports good correlation between Alamar Blue testing and the proportion technique in an earlier study of clinical isolates. This paper’s own study included multidrug-resistant isolates, but compared Alamar Blue results with BACTEC 460, not the conventional proportion method. Its Discussion describes MTT results as preliminary and similar to Alamar Blue results; it does not establish proportion-method correlation for MTT or resazurin as separate assays. Page numbers are unavailable in our copy. (LLM, conf=0.86)
Experimental — accuracy not yet measuredA rapid and inexpensive colorimetric method based on Alamar blue and MTT has been successfully used for determining MICs of first-line drugs in DST of M. tuberculosis.
“A rapid and inexpensive colorimetric method based on the oxidation-reduction indicators Alamar blue and MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium] have been successfully used for determining MICs of first-line drugs in DST of Mycobacterium tuberculosis (1, 9, 20, 31, 38, 49).”
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 [20]
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- Authors
- Franzblau, S. G., Witzig, R. S., McLaughlin, J. C., Torres, P., Madico, G., Hernandez, A., Degnan, M. T., Cook, M. B., Quenzer, V. K., Ferguson, R. M., & Gilman, R. H.
- Affiliations
- <!--label omitted: 1-->GWL Hansen’s Disease Center, Baton Rouge, Louisiana1;
- Johns Hopkins University (US)
- University of New Mexico (US)
- Universidad Peruana Cayetano Heredia (PE)
- <!--label omitted: 3-->University of New Mexico Health Science Center3 and
- New Mexico Department of Health (US)
- Venue
- Journal of Clinical Microbiology
- Publisher
- American Society for Microbiology
- Published
- 1998-02
- Volume
- 36
- Issue
- 2
- Pages
- 362-366
- ISSN
- 0095-1137 (ISSN-L)
- 1098-660X
- PMC
- PMC104543
- OpenAlex
- W2160787060
- Cited by
- 964
- References
- 0
- Citations by year
- 2026: 29 · 2025: 35 · 2024: 42 · 2023: 40 · 2022: 41 · 2021: 42 · 2020: 29 · 2019: 34 · 2018: 44 · 2017: 50 · 2016: 44 · 2015: 59 · 2014: 64 · 2013: 55 · 2012: 45
- FWCI
- 5.29
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Journal of Clinical Microbiology (publishedVersion)
- PubMed Central (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institute of Allergy and Infectious Diseases (N01 AI050016, Y1-AI-50016)
- 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
- Mycobacterium tuberculosis, antimicrobial susceptibility testing, Alamar Blue assay, microplate assay, isoniazid, rifampicin, streptomycin, ethambutol, multidrug-resistant tuberculosis, Peru
- MeSH terms
- Antibiotics, Antitubercular, Antitubercular Agents, Bacteriological Techniques, Culture Media, Drug Resistance, Microbial, Coloring Agents, Ethambutol, Humans, Isoniazid, Microbial Sensitivity Tests, Mycobacterium tuberculosis, Oxazines, Peru, Rifampin, Sensitivity and Specificity, Streptomycin, Tuberculosis, Xanthenes, Drug Resistance, Multiple
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