Interlaboratory study for the evaluation of three microtiter plate-based biofilm quantification methods

• Type: Article in International Scientific Journal
• Date: 2021

Authors

Allkja, J (Author) Other; van Charante, F (Author) Other; Aizawa, J (Author) Other; Reigada, I (Author) Other; Guarch Perez, C (Author) Other; Vazquez Rodriguez, JA (Author) Other; Cos, P (Author) Other; Coenye, T (Author) Other; Fallarero, A (Author) Other; Zaat, SAJ (Author) Other; Felici, A (Author) Other; Ferrari, L (Author) Other; Azevedo, NF (Author) FEUP; Parker, AE (Author) Other; Goeres, DM (Author) Other

Journal

Title: Scientific Reports

Vol. 11

ISSN: 2045-2322

Publisher: Springer Nature

Indexing

ISI Web of Knowledge - 0 Citations

Scopus - 1 Citation

Other information

Authenticus ID: P-00V-4Q1

DOI: 10.1038/s41598-021-93115-w

Abstract (EN): Microtiter plate methods are commonly used for biofilm assessment. However, results obtained with these methods have often been difficult to reproduce. Hence, it is important to obtain a better understanding of the repeatability and reproducibility of these methods. An interlaboratory study was performed in five different laboratories to evaluate the reproducibility and responsiveness of three methods to quantify Staphylococcus aureus biofilm formation in 96-well microtiter plates: crystal violet, resazurin, and plate counts. An inter-lab protocol was developed for the study. The protocol was separated into three steps: biofilm growth, biofilm challenge, biofilm assessment. For control experiments participants performed the growth and assessment steps only. For treatment experiments, all three steps were performed and the efficacy of sodium hypochlorite (NaOCl) in killing S. aureus biofilms was evaluated. In control experiments, on the log(10)-scale, the reproducibility SD (S-R) was 0.44 for crystal violet, 0.53 for resazurin, and 0.92 for the plate counts. In the treatment experiments, plate counts had the best responsiveness to different levels of efficacy and also the best reproducibility with respect to responsiveness (Slope/S-R=1.02), making it the more reliable method to use in an antimicrobial efficacy test. This study showed that the microtiter plate is a versatile and easy-to-use biofilm reactor, which exhibits good repeatability and reproducibility for different types of assessment methods, as long as a suitable experimental design and statistical analysis is applied.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 10