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Interlaboratory study for the evaluation of three microtiter plate-based biofilm quantification methods. / Allkja, Jontana; van Charante, Frits; Aizawa, Juliana et al.

In: Scientific reports, Vol. 11, No. 1, 13779, 01.12.2021.

Research output: Contribution to journalArticleAcademicpeer-review

Harvard

Allkja, J, van Charante, F, Aizawa, J, Reigada, I, Guarch-Pérez, C, Vazquez-Rodriguez, JA, Cos, P, Coenye, T, Fallarero, A, Zaat, SAJ, Felici, A, Ferrari, L, Azevedo, NF, Parker, AE & Goeres, DM 2021, 'Interlaboratory study for the evaluation of three microtiter plate-based biofilm quantification methods', Scientific reports, vol. 11, no. 1, 13779. https://doi.org/10.1038/s41598-021-93115-w

APA

Allkja, J., van Charante, F., Aizawa, J., Reigada, I., Guarch-Pérez, C., Vazquez-Rodriguez, J. A., Cos, P., Coenye, T., Fallarero, A., Zaat, S. A. J., Felici, A., Ferrari, L., Azevedo, N. F., Parker, A. E., & Goeres, D. M. (2021). Interlaboratory study for the evaluation of three microtiter plate-based biofilm quantification methods. Scientific reports, 11(1), [13779]. https://doi.org/10.1038/s41598-021-93115-w

Vancouver

Allkja J, van Charante F, Aizawa J, Reigada I, Guarch-Pérez C, Vazquez-Rodriguez JA et al. Interlaboratory study for the evaluation of three microtiter plate-based biofilm quantification methods. Scientific reports. 2021 Dec 1;11(1):13779. doi: 10.1038/s41598-021-93115-w

Author

Allkja, Jontana ; van Charante, Frits ; Aizawa, Juliana et al. / Interlaboratory study for the evaluation of three microtiter plate-based biofilm quantification methods. In: Scientific reports. 2021 ; Vol. 11, No. 1.

BibTeX

@article{255f5819f41b4aeebd4c6a698493d7f4,
title = "Interlaboratory study for the evaluation of three microtiter plate-based biofilm quantification methods",
abstract = "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 log10-scale, the reproducibility SD (SR) 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/SR = 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.",
author = "Jontana Allkja and {van Charante}, Frits and Juliana Aizawa and In{\'e}s Reigada and Clara Guarch-P{\'e}rez and Vazquez-Rodriguez, {Jesus Augusto} and Paul Cos and Tom Coenye and Adyary Fallarero and Zaat, {Sebastian A. J.} and Antonio Felici and Livia Ferrari and Azevedo, {Nuno F.} and Parker, {Albert E.} and Goeres, {Darla M.}",
note = "Funding Information: This project has received funding from the European Union{\textquoteright}s Horizon 2020 research and innovation program under the Marie Sklodowska—Curie Grant Agreement No. 722467, Print-Aid project. The information and views set out in this article are those of the authors and do not necessarily reflect the official opinion of the European Union. Neither the European Union institutions and bodies nor any person acting on their behalf may be held responsible for the use which may be made of the information contained therein. This work was financially supported by: Base Funding—UIDB/00511/2020 of the Laboratory for Process Engineering, Environment, Biotechnology and Energy—LEPABE—funded by national funds through the FCT/MCTES (PIDDAC). Publisher Copyright: {\textcopyright} 2021, The Author(s).",
year = "2021",
month = dec,
day = "1",
doi = "10.1038/s41598-021-93115-w",
language = "English",
volume = "11",
journal = "Scientific reports",
issn = "2045-2322",
publisher = "Nature Publishing Group",
number = "1",

}

RIS

TY - JOUR

T1 - Interlaboratory study for the evaluation of three microtiter plate-based biofilm quantification methods

AU - Allkja, Jontana

AU - van Charante, Frits

AU - Aizawa, Juliana

AU - Reigada, Inés

AU - Guarch-Pérez, Clara

AU - Vazquez-Rodriguez, Jesus Augusto

AU - Cos, Paul

AU - Coenye, Tom

AU - Fallarero, Adyary

AU - Zaat, Sebastian A. J.

AU - Felici, Antonio

AU - Ferrari, Livia

AU - Azevedo, Nuno F.

AU - Parker, Albert E.

AU - Goeres, Darla M.

N1 - Funding Information: This project has received funding from the European Union’s Horizon 2020 research and innovation program under the Marie Sklodowska—Curie Grant Agreement No. 722467, Print-Aid project. The information and views set out in this article are those of the authors and do not necessarily reflect the official opinion of the European Union. Neither the European Union institutions and bodies nor any person acting on their behalf may be held responsible for the use which may be made of the information contained therein. This work was financially supported by: Base Funding—UIDB/00511/2020 of the Laboratory for Process Engineering, Environment, Biotechnology and Energy—LEPABE—funded by national funds through the FCT/MCTES (PIDDAC). Publisher Copyright: © 2021, The Author(s).

PY - 2021/12/1

Y1 - 2021/12/1

N2 - 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 log10-scale, the reproducibility SD (SR) 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/SR = 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.

AB - 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 log10-scale, the reproducibility SD (SR) 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/SR = 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.

UR - http://www.scopus.com/inward/record.url?scp=85109131612&partnerID=8YFLogxK

U2 - 10.1038/s41598-021-93115-w

DO - 10.1038/s41598-021-93115-w

M3 - Article

C2 - 34215805

VL - 11

JO - Scientific reports

JF - Scientific reports

SN - 2045-2322

IS - 1

M1 - 13779

ER -

ID: 18919420