School of Food Science and Technology, Yangzhou University, Yangzhou, Jiangsu 225127, PR China; Jiangsu key laboratory of zoonoses, Yangzhou, Jiangsu 225009, PR China.
School of Food Science and Technology, Yangzhou University, Yangzhou, Jiangsu 225127, PR China.
Biosens Bioelectron. 2018 Apr 15;102:396-402. doi: 10.1016/j.bios.2017.11.040. Epub 2017 Nov 11.
A novel screen-printed cell-based electrochemical sensor was developed to assess bacterial quorum signaling molecules, N-acylhomoserine lactones (AHLs). Screen-printed carbon electrode (SPCE), which possesses excellent properties such as low-cost, disposable and energy-efficient, was modified with multi-walled carbon nanotubes (MWNTs) to improve electrochemical signals and enhance the sensitivity. Rat basophilic leukemia (RBL-2H3) mast cells encapsulated in alginate/graphene oxide (NaAgl/GO) hydrogel were immobilized on the MWNTs/SPCE to serve as recognition element. Electrochemical impedance spectroscopy (EIS) was employed to record the cell impedance signal as-influenced by Pseudomonas aeruginosa quorum-sensing molecule, N-3-oxododecanoyl homoserine lactone (3OC-HSL). Experimental results show that 3OC-HSL caused a significant decrease in cell viability in a dose dependent manner. The EIS value decreased with concentrations of 3OC-HSL in the range of 0.1-1μM, and the detection limit for 3OC-HSL was calculated to be 0.094μM. These results were confirmed via cell viability, SEM, TEM analysis. Next, the sensor was successfully applied to monitoring the production of AHLs by spoilage bacteria in three different freshwater fish juice samples which efficiently proved the practicability of this cell based method. Therefore, the proposed cell sensor may serve as an innovative and effective approach to the measurement of quorum signaling molecule and thus provides a new avenue for real-time monitoring the spoilage bacteria in freshwater fish production.
一种新型的基于细胞的丝网印刷电化学传感器被开发出来，用于评估细菌群体感应信号分子，N-酰基高丝氨酸内酯（AHLs）。丝网印刷碳电极（SPCE）具有低成本、一次性和节能等优异性能，通过修饰多壁碳纳米管（MWNTs）来提高电化学信号并增强灵敏度。将包封在藻酸盐/氧化石墨烯（NaAgl/GO）水凝胶中的大鼠嗜碱性白血病（RBL-2H3）肥大细胞固定在 MWNTs/SPCE 上作为识别元件。电化学阻抗谱（EIS）用于记录细胞阻抗信号，该信号受铜绿假单胞菌群体感应分子 N-3-氧代十二酰高丝氨酸内酯（3OC-HSL）的影响。实验结果表明，3OC-HSL 以剂量依赖的方式导致细胞活力显著下降。EIS 值随 3OC-HSL 浓度在 0.1-1μM 范围内呈下降趋势，3OC-HSL 的检测限计算为 0.094μM。通过细胞活力、SEM、TEM 分析证实了这些结果。接下来，该传感器成功应用于监测三种不同淡水鱼汁样品中腐败细菌产生的 AHLs，有效地证明了这种基于细胞的方法的实用性。因此，所提出的细胞传感器可作为测量群体感应分子的创新有效方法，从而为实时监测淡水鱼生产中的腐败细菌提供了新途径。
