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利用 QCM 生物传感器通过糖-凝集素相互作用直接且选择性地检测细菌。

Sugar-Lectin Interactions for Direct and Selective Detection of Bacteria Using QCM Biosensor.

机构信息

Chemistry and Environmental Science, New Jersey Institute of Technology, University Heights, Newark, NJ 07102, USA.

Department of Chemistry, Center for Research in Advanced Sensing Technologies & Environmental Sustainability (CREATES), State University of New York at Binghamton, Binghamton, NY 13902, USA.

出版信息

Biosensors (Basel). 2023 Mar 3;13(3):337. doi: 10.3390/bios13030337.

Abstract

Pathogenic () remains a safety concern in the preservation and quality of green leafy vegetables. Sugar-lectin interactions provide a reliable, specific, and effective sensing platform for the detection of bacteria as compared to the tedious conventional plate counting technique. Herein, we present the synthesis of 4-(N-mannosyl) benzoic acid (4-NMBA) and 4-thiophenyl-N-mannose (4-TNM) via a two-step reductive amination for the detection of using a quartz crystal microbalance (QCM) biosensor. The 4-NMBA was synthesized with mannose and para-aminobenzoic (4-PBA), while the 4-TNM was synthesized with mannose and 4-aminophenyl disulfide (4-AHP) using water and acetic acid in a 1:1 ratio. The resultant structure of mannose derivatives (4-NMBA and 4-TNM) was characterized and confirmed using analytical tools, such as Mass Spectrometer, SEM, and FTIR. The choice of ligands (mannose derivatives) is ascribed to the specific recognition of mannose to the FimH lectin of the type 1 pilus of . Furthermore, the 4-PBA and 4-AHP conjugated to mannose increase the ligand affinity to FimH lectins. The setup of the QCM biosensor was composed of modification of the crystal surface and the covalent attachment of ligands for the detection of The piezoelectric effect (frequency shift of the quartz) was proportional to the change in mass added to the gold crystal surface. Both the 4-NMBA- and 4-TNM-coated QCM sensors had a limit of detection of 3.7 CFU/mL and 6.6 CFU/mL with a sensitivity of 2.56 × 10 ng/mL and 8.99 × 10 ng/mL, respectively, within the dynamic range of 10 to 10 CFU/mL. This study demonstrates the application of ligand-coated QCM biosensors as a cost-effective, simple, and label-free technology for monitoring pathogenic bacteria via molecular interactions on crystal surfaces.

摘要

食源性病原体()仍然是绿色叶菜类食品保存和质量的安全隐患。与繁琐的传统平板计数技术相比,糖-凝集素相互作用为细菌检测提供了一种可靠、特异和有效的传感平台。在此,我们通过两步还原胺化法合成了 4-(N-甘露糖基)苯甲酸(4-NMBA)和 4-噻吩基-N-甘露糖(4-TNM),用于使用石英晶体微天平(QCM)生物传感器检测 。4-NMBA 是由甘露糖和对氨基苯甲酸(4-PBA)合成的,而 4-TNM 是由甘露糖和 4-氨基苯基二硫化物(4-AHP)在水和乙酸的 1:1 比例下合成的。甘露糖衍生物(4-NMBA 和 4-TNM)的结构通过质谱仪、SEM 和 FTIR 等分析工具进行了表征和确认。选择配体(甘露糖衍生物)是因为甘露糖能够特异性识别 1 型菌毛的 FimH 凝集素。此外,与甘露糖偶联的 4-PBA 和 4-AHP 增加了配体与 FimH 凝集素的亲和力。QCM 生物传感器的设置由晶体表面的修饰和配体的共价连接组成,用于检测 。石英晶体的压电效应(晶体表面质量增加引起的频率变化)与添加到金晶体表面的质量变化成正比。4-NMBA 和 4-TNM 涂层的 QCM 传感器的检测限分别为 3.7 CFU/mL 和 6.6 CFU/mL,灵敏度分别为 2.56×10 ng/mL 和 8.99×10 ng/mL,动态范围为 10 至 10 CFU/mL。这项研究展示了配体涂层 QCM 生物传感器作为一种具有成本效益、简单且无标记的技术,可通过晶体表面的分子相互作用监测食源性病原体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ed8/10046022/f29fd03a01f4/biosensors-13-00337-sch001.jpg

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