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一种使用重组生物发光来评估细菌黏附的定量方法。

A quantitative method to assess bacterial adhesion using recombinant bioluminescent .

作者信息

Wang Lu, Qiao Xinhua, Gao Lei, Chen Chang, Wan Yi

机构信息

Microbiology Institute of Shaanxi, Xi'an 710043, China.

National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.

出版信息

Biophys Rep. 2021 Feb 28;7(1):55-70. doi: 10.52601/bpr.2021.200043.

Abstract

Bioluminescence technology has been widely used in the field of medical detection. The bioluminescent 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 containing a reporter. By monitoring the growth and bioluminescent properties of the recombinant PAO1- strain, the optimal test conditions for bacterial adhesion detection were determined to be as follows: an initial inoculation density of 10 to 10 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.

摘要

生物发光技术已广泛应用于医学检测领域。生物发光报告系统提供了一个非侵入性平台,可实时监测细菌的生长和表达。本研究旨在建立一种利用含报告基因的重组生物发光菌检测包括医疗器械在内的材料表面细菌黏附的方法。通过监测重组PAO1菌株的生长和生物发光特性,确定细菌黏附检测的最佳试验条件如下:初始接种密度为10⁶至10⁷CFU/mL,pH 6.2的M9培养基,黏附时间为6小时,以及通过超声清洗收集黏附细菌。比较了传统的CFU计数法和生物发光法,并通过测试细菌在各种材料表面的黏附情况验证了该新方法的适用性。经验证的生物发光菌株有望成为细菌黏附评估等应用中的细菌检测工具,以及传统微生物检测程序的补充和替代方法。此外,该方法有可能用于研究无生命物体和活组织表面的细菌黏附。随着该方法的发展及其广泛适用性,有望成为细菌黏附检测和抗黏附材料筛选的标准方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d977/10240536/93e4ec136e55/br-7-1-55-1.jpg

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