Ruggedness and reproducibility of the MBEC biofilm disinfectant efficacy test

• Published in: Journal of microbiological methods 2014-07, Vol.102, p.55-64
• Main Authors: Parker, A.E., Walker, D.K., Goeres, D.M., Allan, N., Olson, M.E., Omar, A.
• Format: Article
• Language: English
• Subjects: Biofilm; Biofilms - drug effects; Disinfectants - pharmacology; Hydrogen-Ion Concentration; Microbial Sensitivity Tests - methods; Pseudomonas aeruginosa; Pseudomonas aeruginosa - drug effects; Pseudomonas aeruginosa - physiology; Reproducibility; Reproducibility of Results; Ruggedness; Sonication; Temperature; Time Factors
• ISSN: 0167-7012; 1872-8359
• DOI: 10.1016/j.mimet.2014.04.013

The MBEC™ Physiology & Genetics Assay recently became the first approved ASTM standardized biofilm disinfectant efficacy test method. This report summarizes the results of the standardization process using Pseudomonas aeruginosa biofilms. Initial ruggedness testing of the MBEC method suggests that the assay is rugged (i.e., insensitive) to small changes to the protocol with respect to 4 factors: incubation time of the bacteria (when varied from 16 to 18h), treatment temperature (20–24°C), sonication duration (25–35min), and sonication power (130–480W). In order to assess the repeatability of MBEC results across multiple tests in the same laboratory and the reproducibility across multiple labs, an 8-lab study was conducted in which 8 concentrations of each of 3 disinfectants (a non-chlorine oxidizer, a phenolic, and a quaternary ammonium compound) were applied to biofilms using the MBEC method. The repeatability and reproducibility of the untreated control biofilms were acceptable, as indicated by small repeatability and reproducibility standard deviations (SD) (0.33 and 0.67 log10(CFU/mm2), respectively). The repeatability SDs of the biofilm log reductions after application of the 24 concentration and disinfectant combinations ranged from 0.22 to 1.61, and the reproducibility SDs ranged from 0.27 to 1.70. In addition, for each of the 3 disinfectant types considered, the assay was statistically significantly responsive to the increasing treatment concentrations. •We outline results for the first standardized biofilm disinfectant efficacy test.•Ruggedness with respect to operational factors was assessed.•Resemblance of controls across multiple laboratories was assessed.•Reproducibility of the biofilm log reductions after disinfection was assessed.•Responsiveness to increasing disinfectant concentrations was assessed.