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Data Quality in Biofilm High-Throughput Routine Analysis: Intralaboratory Protocol Adaptation and Experiment Reproducibility

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Biofilm research is growing more diverse and dependent on high-throughput technologies, and the large-scale production of results aggravates data substantiation. In particular, experimental protocols are often 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 intralaboratory 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 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide inner salt (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. Intralaboratory validation of non-standard protocols is found critical to ensure data quality and enable the comparison of results within and among laboratories.
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Document Type: Research Article

Affiliations: 1: CEB-Centre of Biological Engineering, LIBRO-Laboratory of Research in Biofilms Rosário Oliveira, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal 2: ESEI-Escuela Superior de Ingeniería Informática, Department of Computer Science, University of Vigo, Ourense, Spain; CEB-Centre of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal

Publication date: 01 November 2015

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  • The Journal of AOAC INTERNATIONAL publishes refereed papers and reviews in the fields of chemical, biological and toxicological analytical chemistry for the purpose of showcasing the most precise, accurate and sensitive methods for analysis of foods, food additives, supplements and contaminants, cosmetics, drugs, toxins, hazardous substances, pesticides, feeds, fertilizers and the environment available at that point in time. The scope of the Journal includes unpublished original research describing new analytical methods, techniques and applications; improved approaches to sampling, both in the field and the laboratory; better methods of preparing samples for analysis; collaborative studies substantiating the performance of a given method; statistical techniques for evaluating data. The Journal will also publish other articles of general interest to its audience, e.g., technical communications; cautionary notes; comments on techniques, apparatus, and reagents.
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