T/CI 011-2021 Guidance for culturing, observing, and evaluating antimicrobial resistance in bacterial biofilms

Scope

Scope: This guideline specifies the technical requirements for experimental conditions, experimental design, implementation, and validation of techniques for culturing, observing, and assessing the drug resistance of bacterial biofilms. This guideline is applicable to research institutions, educational institutions, and other biotechnology-related organizations or companies that need to cultivate and observe bacterial biofilms and test their drug resistance. It is also suitable for use by experimental operators in the fields of identifying the biofilm formation ability of clinical and foodborne isolates, determining the drug resistance of these isolates, exploring the molecular mechanisms of biofilm formation and drug resistance, and developing prevention and control methods for bacterial biofilms and related drugs. This guideline can serve as a technical basis for laboratory work involving the cultivation and construction of bacterial biofilms, their observation and quantification, and drug resistance assessments. According to the Patent Law of the People's Republic of China, the use of patented technologies (US9988380B2, US10416153B2, ZL201711231930.8, CN110974838A) mentioned in this guideline requires written permission from the patent owner. Main technical content: Methods and technical standards for cultivating bacterial biofilms. The following biofilm cultivation methods and technical standards are written using Pseudomonas aeruginosa as a model strain. Cultivation and experimental conditions for other strains can be adjusted as needed to achieve optimal experimental results. Pseudomonas aeruginosa (PA) is a common Gram-negative opportunistic pathogen that is widely distributed in normal human skin, respiratory tracts, intestines, and various natural environments such as water and soil. This bacterium has a rod-shaped, single flagellum morphology and an optimal growth temperature of 25-37°C, although it can also grow at 42°C. On LB medium, it forms green-b

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