Autor(es):
Jorge, Paula Alexandra Silva ; Lourenço, Anália ; Pereira, Maria Olívia
Data: 2015
Identificador Persistente: http://hdl.handle.net/1822/39597
Origem: RepositóriUM - Universidade do Minho
Projeto/bolsa:
info:eu-repo/grantAgreement/FCT/5876/147337/PT;
info:eu-repo/grantAgreement/FCT/5876-PPCDTI/113196/PT
;
info:eu-repo/grantAgreement/FCT/5876-PPCDTI/126270/PT
;
info:eu-repo/grantAgreement/EC/FP7/316265/EU;
info:eu-repo/grantAgreement/FCT/COMPETE/126270/PT;
Assunto(s): Science & Technology
Descrição
Biofilm research is growing more diverse and dependent on high-throughput technologies and the large-scale production of results aggravates data substantiation. In particular, it is often the case that experimental protocols are adapted to meet the needs of a particular laboratory and no statistical validation of the modified method is provided. This paper discusses the impact of intra-laboratory adaptation and non-rigorous documentation of experimental protocols on biofilm data interchange and validation. The case study is a non-standard, but widely used, workflow for Pseudomonas aeruginosa biofilm development, considering three analysis assays: the crystal violet (CV) assay for biomass quantification, the XTT assay for respiratory activity assessment, and the colony forming units (CFU) assay for determination of cell viability. The ruggedness of the protocol was assessed by introducing small changes in the biofilm growth conditions, which simulate minor protocol adaptations and non-rigorous protocol documentation. Results show that even minor variations in the biofilm growth conditions may affect the results considerably, and that the biofilm analysis assays lack repeatability. Intra-laboratory validation of non-standard protocols is found critical to ensure data quality and enable the comparison of results within and among laboratories.
We thank the Portuguese Foundation for Science and Technology (FCT) the strategic funding of UID/BIO/04469/2013 unit. The authors also acknowledge the support by FCT and the European Community fund FEDER, through Program COMPETE, under the scope of the Projects AntiPep PTDC/SAU-SAP/113196/2009 (FCOMP-01-0124-FEDER-016012), RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462), the Projects "BioHealth-Biotechnology and Bioengineering approaches to improve health quality," NORTE-07-0124-FEDER-000027, co-funded by the Programa Operacional Regional do Norte (ON.2-O Novo Norte), QREN, FEDER, and the Agrupamento INBIOMED from DXPCTSUG-FEDER unha maneira de facer Europa (2012/273). The research leading to these results has received funding from the European Union's Seventh Framework Programme FP7/REGPOT-2012-2013.1 under grant agreement No. 316265, BIOCAPS. This document reflects only the author's views, and the European Union is not liable for any use that may be made of the information contained herein. The authors also acknowledge the Ph.D. Grant of Paula Jorge, Ref. SFRH/BD/88192/2012.
