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...

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Autores principales:	Jontana Allkja, Frits van Charante, Juliana Aizawa, Inés Reigada, Clara Guarch-Pérez, Jesus Augusto Vazquez-Rodriguez, Paul Cos, Tom Coenye, Adyary Fallarero, Sebastian A. J. Zaat, Antonio Felici, Livia Ferrari, Nuno F. Azevedo, Albert E. Parker, Darla M. Goeres
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e846ad0bf22d451e93a5a430e7a0ebe2
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spelling	oai:doaj.org-article:e846ad0bf22d451e93a5a430e7a0ebe22021-12-02T14:33:51ZInterlaboratory study for the evaluation of three microtiter plate-based biofilm quantification methods10.1038/s41598-021-93115-w2045-2322https://doaj.org/article/e846ad0bf22d451e93a5a430e7a0ebe22021-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-93115-whttps://doaj.org/toc/2045-2322Abstract 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.Jontana AllkjaFrits van CharanteJuliana AizawaInés ReigadaClara Guarch-PérezJesus Augusto Vazquez-RodriguezPaul CosTom CoenyeAdyary FallareroSebastian A. J. ZaatAntonio FeliciLivia FerrariNuno F. AzevedoAlbert E. ParkerDarla M. GoeresNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jontana Allkja Frits van Charante Juliana Aizawa Inés Reigada Clara Guarch-Pérez Jesus Augusto Vazquez-Rodriguez Paul Cos Tom Coenye Adyary Fallarero Sebastian A. J. Zaat Antonio Felici Livia Ferrari Nuno F. Azevedo Albert E. Parker Darla M. Goeres Interlaboratory study for the evaluation of three microtiter plate-based biofilm quantification methods
description	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.
format	article
author	Jontana Allkja Frits van Charante Juliana Aizawa Inés Reigada Clara Guarch-Pérez Jesus Augusto Vazquez-Rodriguez Paul Cos Tom Coenye Adyary Fallarero Sebastian A. J. Zaat Antonio Felici Livia Ferrari Nuno F. Azevedo Albert E. Parker Darla M. Goeres
author_facet	Jontana Allkja Frits van Charante Juliana Aizawa Inés Reigada Clara Guarch-Pérez Jesus Augusto Vazquez-Rodriguez Paul Cos Tom Coenye Adyary Fallarero Sebastian A. J. Zaat Antonio Felici Livia Ferrari Nuno F. Azevedo Albert E. Parker Darla M. Goeres
author_sort	Jontana Allkja
title	Interlaboratory study for the evaluation of three microtiter plate-based biofilm quantification methods
title_short	Interlaboratory study for the evaluation of three microtiter plate-based biofilm quantification methods
title_full	Interlaboratory study for the evaluation of three microtiter plate-based biofilm quantification methods
title_fullStr	Interlaboratory study for the evaluation of three microtiter plate-based biofilm quantification methods
title_full_unstemmed	Interlaboratory study for the evaluation of three microtiter plate-based biofilm quantification methods
title_sort	interlaboratory study for the evaluation of three microtiter plate-based biofilm quantification methods
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/e846ad0bf22d451e93a5a430e7a0ebe2
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Interlaboratory study for the evaluation of three microtiter plate-based biofilm quantification methods

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