High-throughput combination assay for studying biofilm formation of uropathogenic Escherichia coli.
Arch Microbiol
; 206(8): 344, 2024 Jul 05.
Article
em En
| MEDLINE
| ID: mdl-38967798
ABSTRACT
Uropathogenic Escherichia coli, the most common cause for urinary tract infections, forms biofilm enhancing its antibiotic resistance. To assess the effects of compounds on biofilm formation of uropathogenic Escherichia coli UMN026 strain, a high-throughput combination assay using resazurin followed by crystal violet staining was optimized for 384-well microplate. Optimized assay parameters included, for example, resazurin and crystal violet concentrations, and incubation time for readouts. For the assay validation, quality parameters Z' factor, coefficient of variation, signal-to-noise, and signal-to-background were calculated. Microplate uniformity, signal variability, edge well effects, and fold shift were also assessed. Finally, a screening with known antibacterial compounds was conducted to evaluate the assay performance. The best conditions found were achieved by using 12 µg/mL resazurin for 150 min and 0.023% crystal violet. This assay was able to detect compounds displaying antibiofilm activity against UMN026 strain at sub-inhibitory concentrations, in terms of metabolic activity and/or biomass.
Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Oxazinas
/
Xantenos
/
Biofilmes
/
Escherichia coli Uropatogênica
/
Ensaios de Triagem em Larga Escala
/
Violeta Genciana
/
Antibacterianos
Limite:
Humans
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article