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基于阵列的微生物鉴定方法,用于检测细胞外氨基糖苷类抗生素残留。

Array-based microbial identification upon extracellular aminoglycoside residue sensing.

机构信息

Key Laboratory of Biomedical Information Engineering of Ministry of Education, Institute of Analytical Chemistry and Instrument for Life Science, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, 710049, Shaanxi, China.

Department of Chemical Engineering, The University of Western Australia, Perth, WA, 6009, Australia.

出版信息

Anal Bioanal Chem. 2021 Jul;413(18):4689-4696. doi: 10.1007/s00216-021-03346-z. Epub 2021 Apr 23.

Abstract

Sensitive and rapid identification of pathogenic microorganisms is of great importance for clinical diagnosis and treatment. In this study, we developed an ultrasensitive colorimetric sensor array (CSA) based on the interactions between aminoglycoside antibiotics (AMGs) and Ag nanoparticles decorated with β-cyclodextrin (AgNPs@β-CD) to discriminate microorganisms quickly and accurately. Microorganisms can absorb different amounts of AMGs after incubation. Upon the addition of AgNPs@β-CD, the corresponding extracellular AMG residues will bind to AgNPs@β-CD, leading to color changes due to the modifications in localized surface plasmon resonance. The array was developed using 4 AMGs as sensing elements and AgNPs@β-CD as the colorimetric probe to generate a unique colorimetric response pattern for each microorganism. Standard chemometric methods indicated excellent discrimination among 20 microorganisms at low concentrations of 2 × 10 CFU/mL. Therefore, this ultrasensitive CSA can be used for microbial discrimination portably and efficiently. Importantly, the concentration of microbial discrimination by our array is much lower than that of prior CSAs. This method of extracellular residue sensing also provided a new strategy to improve the sensitivity of conventional CSA in the discrimination of microorganisms, to measure the amount of intercellular uptake of AMGs by microorganisms, and to screen drugs that can easily be accumulated by the pathogenic microorganisms.

摘要

快速灵敏地识别致病微生物对于临床诊断和治疗至关重要。在本研究中,我们开发了一种基于氨基糖苷类抗生素(AMGs)与β-环糊精(β-CD)修饰的银纳米颗粒(AgNPs@β-CD)相互作用的超灵敏比色传感器阵列(CSA),可快速准确地识别微生物。微生物在孵育后可以吸收不同量的 AMGs。加入 AgNPs@β-CD 后,相应的细胞外 AMG 残基将与 AgNPs@β-CD 结合,由于局部表面等离子体共振的变化,导致颜色发生变化。该阵列使用 4 种 AMGs 作为传感元件,AgNPs@β-CD 作为比色探针,为每种微生物生成独特的比色响应模式。标准化学计量学方法表明,在 2×10^CFU/mL 的低浓度下,可对 20 种微生物进行出色的区分。因此,这种超灵敏 CSA 可用于便携且高效地进行微生物鉴别。重要的是,我们的阵列对微生物的区分浓度远低于先前的 CSA。这种细胞外残留感应方法还为提高传统 CSA 在微生物鉴别中的灵敏度提供了一种新策略,可用于测量微生物摄取细胞内 AMGs 的量,以及筛选易于被致病菌积累的药物。

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