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基于新型肌尾噬菌体 SEP37 的 EIS 生物传感器,用于快速、特异性检测食品基质中的沙门氏菌。

EIS biosensor based on a novel Myoviridae bacteriophage SEP37 for rapid and specific detection of Salmonella in food matrixes.

机构信息

College of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.

Faculty of Science, Huazhong Agricultural University, Wuhan 430070, China.

出版信息

Food Res Int. 2022 Aug;158:111479. doi: 10.1016/j.foodres.2022.111479. Epub 2022 Jun 17.

Abstract

Recently, using bacteriophages as new molecular probes in reliable platforms for the detection of bacterial pathogens has attracted more and more increasing attentions. In this paper, a novel isolated Myoviridae bacteriophage SEP37 was covalently immobilized onto gold nanoparticles (AuNPs) modified gold disk electrode (GDE) surfaces using cysteamine (Cys) as a crosslinker. Substrates of GDE-AuNPs-Cys-Phage SEP37 and specific capture of Salmonella cells had been characterized using scanning electron microscopy (SEM) separately. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used to study the electrochemical response of the biosensor interface manufacturing and bacterial capture process. Under the optimal experimental conditions, this phage-based EIS biosensor was able to detect Salmonella with a wide linear range from 2 × 10 to 2 × 10 colony forming unit (CFU)/mL within 30 min in spiked lake water and lettuce samples, with a limit of detection (LOD) of 17 CFU/mL. The detection linear range of spiked chicken samples was 2 × 10 to 2 × 10 CFU/mL, with a LOD of 1.3 × 10 CFU/mL. In combination with a pre-enrichment process for 3.5 h, this assay could reach a LOD of 1 CFU/mL in chicken breast meat samples. Besides, this phage-based EIS biosensor provided good reproducibility and stability. This phage-based EIS biosensor opens a new opportunity for the detection of pathogenic bacteria using the inherent selectivity of bacteriophage recognition.

摘要

最近,利用噬菌体作为新的分子探针,在可靠的平台上检测细菌病原体,引起了越来越多的关注。在本文中,一种新型的分离自肌尾噬菌体科的噬菌体 SEP37 被共价固定在金纳米粒子(AuNPs)修饰的金盘电极(GDE)表面上,使用半胱氨酸(Cys)作为交联剂。GDE-AuNPs-Cys-Phage SEP37 的底物和沙门氏菌细胞的特异性捕获分别通过扫描电子显微镜(SEM)进行了表征。循环伏安法(CV)和电化学阻抗谱(EIS)用于研究生物传感器界面制造和细菌捕获过程的电化学响应。在最佳实验条件下,该基于噬菌体的 EIS 生物传感器能够在 30 分钟内检测到浓度范围为 2×10 至 2×10 个菌落形成单位(CFU)/mL 的湖水和生菜样本中的沙门氏菌,检测限(LOD)为 17 CFU/mL。在加标鸡肉样本中的检测线性范围为 2×10 至 2×10 CFU/mL,检测限(LOD)为 1.3×10 CFU/mL。与 3.5 小时的预富集过程相结合,该检测方法可以达到鸡肉样本中 1 CFU/mL 的检测限。此外,该基于噬菌体的 EIS 生物传感器具有良好的重现性和稳定性。该基于噬菌体的 EIS 生物传感器为利用噬菌体识别的固有选择性检测致病菌开辟了新的机会。

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