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Microplate alamarBlue assay for Paracoccidioides susceptibility testing
A C A de Paula e Silva1, H C Oliveira, J F Silva
1Faculdade de Ciências Farmacêuticas, UNESP-Universidade Estadual Paulista, Araraquara, Departamento de Analises Clinicas, Laboratório de Micologia Clínica, São Paulo, Brazil.
Journal of Clinical Microbiology
|January 25, 2013
Summary
A new microdilution method using alamarBlue was developed for dimorphic fungi like Paracoccidioides. This sensitive and inexpensive test effectively monitors antifungal drug activity in slow-growing microorganisms.
Area of Science:
- Medical Mycology
- Antimicrobial Susceptibility Testing
Background:
- The Clinical and Laboratory Standards Institute (CLSI) M27-A3 method is unsuitable for dimorphic fungi, including the Paracoccidioides genus.
- Accurate antifungal susceptibility testing is crucial for treating infections caused by Paracoccidioides species.
Purpose of the Study:
- To develop and validate a novel microdilution method for assessing the susceptibility of Paracoccidioides brasiliensis and Paracoccidioides lutzii to antifungal agents.
- To incorporate the alamarBlue reagent for a practical and sensitive detection system.
Main Methods:
- Development of a modified microdilution assay.
- Inclusion of the alamarBlue reagent for colorimetric detection of fungal growth and viability.
- Testing of Paracoccidioides brasiliensis and Paracoccidioides lutzii against amphotericin B and itraconazole.
Main Results:
- The developed microdilution method demonstrated sensitivity in detecting fungal responses.
- The assay proved practical and cost-effective for evaluating antifungal activity.
- The method is suitable for monitoring low-growth microorganisms like Paracoccidioides.
Conclusions:
- A novel, sensitive, practical, and inexpensive microdilution method using alamarBlue has been established for Paracoccidioides species.
- This method facilitates the monitoring of antifungal drug activity against challenging dimorphic fungi.
- The findings support the clinical utility of this assay for guiding treatment decisions.
