Whole-Cell Pseudomonas aeruginosa Localized Surface Plasmon Resonance Aptasensor.
Whole-Cell Pseudomonas aeruginosa Localized Surface Plasmon Resonance Aptasensor.
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DOI：
10.1021/acs.analchem.7b04800
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发表时间：
2018-02-06
影响因子：
7.4
通讯作者：
Bohn PW
中科院分区：
文献类型：
--
作者：
Hu J;Fu K;Bohn PW
全细胞铜绿假单胞菌的检测提出了一个有趣的挑战，直接应用于医疗保健和预防医院感染。为了解决这一问题，开发了基于局部表面等离子体共振（LSPR）的传感平台，以使用表面限定的适体作为亲和试剂来检测全细胞铜绿假单胞菌菌株PAO 1。纳米球光刻（NSL）被用来制造传感器表面包含一个六边形阵列的Au纳米三角形。随后用生物素化的聚乙二醇（Bt-PEG）硫醇/PEG硫醇（1：3）、中性抗生物素蛋白和生物素化的适体以夹心形式修饰传感器表面。特别选择Bt-PEG硫醇/PEG硫醇的1：3（v/v）比率以最大化PAO 1结合，同时最小化非特异性吸附和空间位阻。与之前的全细胞LSPR工作相反，LSPR波长偏移显示在10-103 cfu mL−1的范围内与细菌浓度线性相关。该LSPR传感平台快速（检测时间约为3小时）、灵敏（低至单个细菌的水平）、对假单胞菌菌株PAO 1的检测选择性优于其他菌株，并且在环境条件下储存时表现出临床相关的动态范围和优异的保质期（≥2个月）。这种多功能的LSPR传感平台应该可以通过切换亲和试剂扩展到广泛的超分子分析物，包括细菌和病毒，并且它具有用于即时护理和现场应用的潜力。
The detection of whole-cell Pseudomonas aeruginosa presents an intriguing challenge with direct applications in health care and the prevention of nosocomial infection. To address this problem, a localized surface plasmon resonance (LSPR) based sensing platform was developed to detect whole-cell Pseudomonas aeruginosa strain PAO1 using a surface-confined aptamer as an affinity reagent. Nanosphere lithography (NSL) was used to fabricate a sensor surface containing a hexagonal array of Au nanotriangles. The sensor surface was subsequently modified with biotinylated polyethylene glycol (Bt-PEG) thiol/PEG thiol (1:3), neutravidin, and biotinylated aptamer in a sandwich format. The 1:3 (v/v) ratio of Bt-PEG thiol/PEG thiol was specifically chosen to maximize PAO1 binding while minimizing nonspecific adsorption and steric hindrance. In contrast to prior whole-cell LSPR work, the LSPR wavelength shift was shown to be linearly related to bacterial concentration over the range of 10–103 cfu mL−1. This LSPR sensing platform is rapid (~3 h for detection), sensitive (down to the level of a single bacterium), selective for detection of Pseudomonas strain PAO1 over other strains, and exhibits a clinically relevant dynamic range and excellent shelf life (≥2 months) when stored at ambient conditions. This versatile LSPR sensing platform should be extendable to a wide range of supermolecular analytes, including both bacteria and viruses, by switching affinity reagents, and it has potential to be used in point-of-care and field-based applications.
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8.4
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