Validation of a modified fluorimetric assay for the screening of trichomonacidal drugs
Mem. Inst. Oswaldo Cruz
;
107(5): 637-643, Aug. 2012. ilus, tab
Article
Dans Anglais
| LILACS
| ID: lil-643749
ABSTRACT
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.
Texte intégral:
Disponible
Indice:
LILAS (Amériques)
Sujet Principal:
Trichomonas vaginalis
/
Clotrimazole
/
Métronidazole
/
Antitrichomonas
Type d'étude:
Etude diagnostique
/
Étude de dépistage
langue:
Anglais
Texte intégral:
Mem. Inst. Oswaldo Cruz
Thème du journal:
Médecine tropicale
/
Parasitologie
Année:
2012
Type:
Article
/
descriptif de projet
Pays d'affiliation:
Cuba
/
Espagne
Institution/Pays d'affiliation:
Central University of Las Villas/CU
/
Universidad Complutense de Madrid/ES
/
Universidad Politécnica de Madrid/ES
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