Validation of a modified fluorimetric assay for the screening of trichomonacidal drugs
AUTOR(ES)
Escribano, Alexandra Ibáñez, Marcel, Alfredo Meneses, Tugores, Yanetsy Machado, Ruiz, Juan José Nogal, Redó, Vicente J Arán, García-Trevijano, José Antonio Escario, Barrio, Alicia Gómez
FONTE
Memórias do Instituto Oswaldo Cruz
DATA DE PUBLICAÇÃO
2012-08
RESUMO
A fluorimetric microassay that uses a redox dye to determine the viability of the flagellate Trichomonas vaginalis has been optimised to provide a more sensitive method to evaluate potential trichomonacidal compounds. Resazurin has been used in recent years to test drugs against different parasites, including trichomonadid protozoa; however, the reproducibility of these resazurin-based methods in our laboratory has been limited because the flagellate culture medium spontaneously reduces the resazurin. The objective of this work was to refine the fluorimetric microassay method previously developed by other research groups to reduce the fluorescence background generated by the media and increase the sensitivity of the screening assay. The experimental conditions, time of incubation, resazurin concentration and media used in the microtitre plates were adjusted. Different drug sensitivity studies against T. vaginalis were developed using the 5-nitroimidazole reference drugs, new 5-nitroindazolinones and 5-nitroindazole synthetic derivatives. Haemocytometer count results were compared with the resazurin assay using a 10% solution of 3 mM resazurin dissolved in phosphate buffered saline with glucose (1 mg/mL). The fluorimetric assay and the haemocytometer counts resulted in similar percentages of trichomonacidal activity in all the experiments, demonstrating that the fluorimetric microtitre assay has the necessary accuracy for high-throughput screening of new drugs against T. vaginalis.
Documentos Relacionados
- A modified fluorimetric procedure for the rapid estimation of oxytetracycline in blood.
- Modified Colorimetric Assay for Susceptibility Testing of Azole Antifungal Drugs against Candida Species
- Microculture screening assay for primary in vitro evaluation of drugs against Pneumocystis carinii.
- Simple Fibroblast-Based Assay for Screening of New Antimicrobial Drugs against Mycobacterium tuberculosis
- Standardization of a fluorimetric assay for the determination of tissue angiotensin-converting enzyme activity in rats