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使用固定化抗菌肽对细菌病原体进行快速电阻抗检测。

Rapid, electrical impedance detection of bacterial pathogens using immobilized antimicrobial peptides.

作者信息

Lillehoj Peter B, Kaplan Christopher W, He Jian, Shi Wenyuan, Ho Chih-Ming

机构信息

1Department of Mechanical Engineering, Michigan State University, East Lansing, MI, USA.

出版信息

J Lab Autom. 2014 Feb;19(1):42-9. doi: 10.1177/2211068213495207. Epub 2013 Jul 12.

Abstract

The detection of bacterial pathogens plays an important role in many biomedical applications, including clinical diagnostics, food and water safety, and biosecurity. Most current bacterial detection technologies, however, are unsuitable for use in resource-limited settings where the highest disease burdens often exist. Thus, there is an urgent need to develop portable, user-friendly biosensors capable of rapid detection of multiple pathogens in situ. We report a microfluidic chip for multiplexed detection of bacterial cells that uses antimicrobial peptides (AMPs) with species-specific targeting and binding capabilities. The AMPs are immobilized onto an electrical impedance microsensor array and serve as biorecognition elements for bacterial cell detection. Characterization of peptide immobilization on the sensors revealed robust surface binding via cysteine-gold interactions and vertical alignment relative to the sensor surface. Samples containing Streptococcus mutans and Pseudomonas aeruginosa were loaded in the chip, and both microorganisms were detected at minimum concentrations of 10⁵ cfu/mL within 25 min. Measurements performed in a variety of solutions revealed that high-conductivity solutions produced the largest impedance values. By integrating a highly specific bacterial cell capture scheme with rapid electrical detection, this device demonstrates great potential as a next-generation, point-of-care diagnostic platform for the detection of disease-causing pathogenic agents.

摘要

细菌病原体的检测在许多生物医学应用中发挥着重要作用,包括临床诊断、食品和水安全以及生物安全。然而,目前大多数细菌检测技术不适用于疾病负担往往最重的资源有限环境。因此,迫切需要开发能够在原位快速检测多种病原体的便携式、用户友好型生物传感器。我们报告了一种用于多重检测细菌细胞的微流控芯片,该芯片使用具有物种特异性靶向和结合能力的抗菌肽(AMPs)。这些抗菌肽被固定在一个电阻抗微传感器阵列上,并作为细菌细胞检测的生物识别元件。对传感器上肽固定化的表征显示,通过半胱氨酸-金相互作用实现了牢固的表面结合,并且相对于传感器表面呈垂直排列。将含有变形链球菌和铜绿假单胞菌的样品加载到芯片中,在25分钟内以最低10⁵ cfu/mL的浓度检测到了这两种微生物。在各种溶液中进行的测量表明,高电导率溶液产生的阻抗值最大。通过将高度特异性的细菌细胞捕获方案与快速电检测相结合,该设备作为检测致病病原体的下一代即时诊断平台显示出巨大潜力。

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