A quantitative method to assess bacterial adhesion using recombinant bioluminescent Pseudomonas aeruginosa

Abstract

Bioluminescence technology has been widely used in the field of medical detection. The bioluminescent lux reporter system provides a non-invasive platform to monitor bacterial growth and expression in real time. This study aimed to establish a method for detecting bacterial adhesion on the surface of materials, including medical devices, by using recombinant bioluminescent Pseudomonas aeruginosa containing a lux reporter. By monitoring the growth and bioluminescent properties of the recombinant PAO1-lux strain, the optimal test conditions for bacterial adhesion detection in vitro were determined to be as follows: an initial inoculation density of 10⁵ to 10⁶ CFU/mL, M9 medium at a pH 6.2, an adhesion time of 6 h, and the collection of adherent bacteria by ultrasonic cleaning. The traditional CFU counting method and the bioluminescence method were compared, and the applicability of the new method was verified by testing the adhesion of bacteria on the surface of various materials. The validated bioluminescent strains could serve as strong candidates to be used as bacterial detection tools in applications such as bacterial adhesion evaluation as well as supplements and alternatives to traditional microbiological testing procedures. In addition, this method has the potential to enable the study of bacterial adhesion on the surface of inanimate objects and living tissues. With the development of this method and its wide applicability, it is expected to become a standard method for the detection of bacterial adhesion and the screening of anti-adhesion materials.

Graphical abstract

Keywords

Bacterial adhesion / PAO1-lux / Bioluminescence assay / Medical materials

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Acknowledgements

This work was supported by the Shaanxi Academy of Sciences Project (2020k-26, 2018nk-01), the Foundation of Science and Technology in Shaanxi Province (2020TD-050), and the National Natural Science Foundation of China (31900893).

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Conflict of interest Lu Wang, Xinhua Qiao, Lei Gao, Chang Chen and Yi Wan declare that they have no conflict of interest. Human and animal rights and informed consent This article does not contain any studies with human or animal subjects performed by any of the authors. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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