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- Title
EVALUATION OF THE ABILITY OF SOME MULTIDRUGRESISTANT BACTERIAL STRAINS TO FORM BIOFILMS.
- Authors
GAȘPAR, CRISTINA MIRABELA; KIS, BRIGITTA; TATU, CARMEN; PANAITESCU, CARMEN; ZAMBORI, CSILLA; SIMINA, ALINA GEORGIANA; TULCAN, CAMELIA; HUȚU, IOAN; VLAD, DALIBORCA CRISTINA; PĂUNESCU, VIRGIL; TĂNASIE, GABRIELA
- Abstract
The crystal violet (CV) based microtitre plate assay is one of the most widely used method in screening for biofilm forming bacterial strains or in evaluating the efficacy of antibiofilm products. Despite the modifications made over the time in order to increase CV staining method's accuracy, this assay still shares the issue of "edge effect", phenomenon due mainly to evaporation of the liquid from the peripheral wells. This allows the planktonic cells to stick to the walls, which in turn binds the CV dye and gives a false result as biofilm biomass, thus increasing the rate of experimental errors. The present study aimed to improve the accuracy of the crystal violet staining assay, by reducing the "edge effect" through certain modifications that involved either changing the incubation conditions, by using a climate chamber instead of a classic incubator or the working technique, by filling the peripheral wells from lines A, B, G, H and columns 1, 2, 11 and 12 of a 96 well microtiter plate with sterile distiled water. Results showed that by filling the peripheral wells with water, the „edge effect" is reduced and the OD values are relatively homogene with lower standard deviations, when compared with incubating in a classic incubator and in the climate chamber (p< .05). Therefore, this improvement to crystal violet based microtiter plate assay has shown to give minimum variability in our results, mainly when the biofilm assay requires long incubation time involving more than 24 h.
- Subjects
GENTIAN violet; STAINS &; staining (Microscopy); BIOFILMS; MICROPLATES; IRON &; steel plates
- Publication
Physiology / Fiziologia, 2021, Vol 31, Issue 2, p11
- ISSN
1223-2076
- Publication type
Article