Biotechnical Faculty, Department of Food Science and Technology, University of Ljubljana, Ljubljana, Slovenia.
Biotechnical Faculty, Department of Microbiology, University of Ljubljana, Ljubljana, Slovenia.
Biosens Bioelectron. 2022 Sep 15;212:114439. doi: 10.1016/j.bios.2022.114439. Epub 2022 May 30.
The food-borne pathogen Campylobacter jejuni produces autoinducer-2 (AI-2) as an interspecies signalling molecule. AI-2 can trigger enhanced colonisation and biofilm formation, and this poses a serious risk to public health. To date, this communication system of C. jejuni is only partially understood, as detection and quantification of such autoinducer signalling molecules in complex media is hard to achieve. We have developed a whole-cell Vibrioharveyi-based biosensor assay to accurately quantify and follow production of AI-2 by C. jejuni 81-176 in a defined growth medium and in a model food system. Several V. harveyi strains were tested, but the most sensitive bioluminescent response to C. jejuni AI-2 was achieved with V. harveyi MM30, likely due to its ability to self-amplify the response to AI-2. The AI-2 concentrations measured by this biosensor were confirmed using an independent analytical method, HPLC-FLD, which we introduced for Campylobacter analytics for the first time. The AI-2 concentration produced by C. jejuni 81-176 in the model food system was ∼5-fold that in the defined growth medium, at the same cell density. Together with the linear increments in AI-2 concentrations with cell density, this suggests that in C. jejuni, AI-2 represents a metabolic by-product rather than a true quorum-sensing molecule. This biosensor method is highly sensitive, as shown by the reduction in the limit of detection (by a factor of 100) compared to HPLC-FLD, and it enables quantification of AI-2 in complex matrices, such as food, which will help to improve the quality and safety of food production.
食源性病原体空肠弯曲菌产生群体感应-2(AI-2)作为一种种间信号分子。AI-2 可以触发增强的定植和生物膜形成,这对公共健康构成严重威胁。迄今为止,这种空肠弯曲菌的通讯系统仅部分被理解,因为在复杂培养基中检测和定量这种自动诱导信号分子是困难的。我们开发了一种基于全细胞 Vibrio harveyi 的生物传感器测定法,以准确地定量和跟踪空肠弯曲菌 81-176 在限定生长培养基和模型食品系统中的 AI-2 产生。测试了几种 V. harveyi 菌株,但对 C. jejuni AI-2 最敏感的生物发光反应是由 V. harveyi MM30 实现的,这可能是由于其自我放大对 AI-2 的反应的能力。通过我们首次引入用于弯曲杆菌分析的 HPLC-FLD 等独立分析方法,证实了该生物传感器测量的 AI-2 浓度。在相同细胞密度下,空肠弯曲菌 81-176 在模型食品系统中产生的 AI-2 浓度约为限定生长培养基中的 5 倍。与细胞密度的 AI-2 浓度线性递增相结合,这表明在空肠弯曲菌中,AI-2 代表一种代谢副产物而不是真正的群体感应分子。这种生物传感器方法非常灵敏,与 HPLC-FLD 相比,检测限降低了一个数量级(降低了 100 倍),并且能够在复杂基质(如食品)中定量 AI-2,这将有助于提高食品生产的质量和安全性。