Albrich Werner C, Kahlert Christian R, Nigg Susanne, Boesel Luciano F, Giovannini Giorgia
Division of Infectious Diseases, Infection Prevention and Travel Medicine, Kantonsspital St. Gallen, Kantonsspital St. Gallen, Rorschacher Strasse 95, St. Gallen 9007, Switzerland.
Infectious Diseases and Hospital Epidemiology, Children's Hospital St. Gallen, Claudiusstr. 6, St. Gallen 9006, Switzerland.
Anal Chem. 2024 Dec 31;96(52):20578-20586. doi: 10.1021/acs.analchem.4c05182. Epub 2024 Dec 16.
Urease-producing bacteria are highly relevant in medicine due to their role in various pathogenic processes and their impact on human health, causing serious medical conditions such as peptic ulcer disease, gastric cancer, and respiratory and urinary tract infections. In this work, we designed fluorescent polymeric particles (PNP_FITC) to enable the detection of urease-producing bacteria by targeting the enzymatic activity of urease. In particular, the PNP_FITC matrix is degraded by urease, leading to a measurable increase in the intensity of the fluorescent signal. This approach is designed to directly sense urease activity and is therefore not affected by environmental parameters, unlike standard methods based on the quantification of enzymatic metabolites (i.e., NH and CO). PNP_FITC exhibited a linear response in the urease range of 0-7.5 U/mL, with a calculated limit of detection of 0.4 U/mL. The direct detection of enzymatic activity makes PNP_FITC suitable for detecting urease-producing bacteria ( and ) with a detection limit of 10 ∧ 3 bacteria/mL, which were not detectable using the pH-based method employed as the reference in this work. Given the improvements achieved with PNP_FITC in terms of robustness, sensitivity, and selectivity of urease detection compared to the standard methods, this approach represents a step forward toward the development of advanced point-of-care, enabling the prompt diagnosis of bacterial infections.
产脲酶细菌在医学领域具有高度相关性，因为它们在各种致病过程中发挥作用，并对人类健康产生影响，可引发诸如消化性溃疡病、胃癌以及呼吸道和泌尿道感染等严重病症。在这项工作中，我们设计了荧光聚合物颗粒（PNP_FITC），通过靶向脲酶的酶活性来实现对产脲酶细菌的检测。具体而言，PNP_FITC基质会被脲酶降解，导致荧光信号强度出现可测量的增加。这种方法旨在直接感知脲酶活性，因此不像基于酶代谢产物（即NH和CO）定量的标准方法那样受环境参数影响。PNP_FITC在0 - 7.5 U/mL的脲酶范围内呈现线性响应，计算得出的检测限为0.4 U/mL。对酶活性的直接检测使得PNP_FITC适用于检测产脲酶细菌（ 和 ），检测限为10∧3个细菌/mL，而使用本工作中用作参考的基于pH值的方法则无法检测到。鉴于与标准方法相比，PNP_FITC在脲酶检测的稳健性、灵敏度和选择性方面取得了改进，这种方法代表了朝着开发先进的即时检测技术迈出的一步，能够实现细菌感染的快速诊断。
