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基于石英晶体微天平主动和被动模式的微生物快速检测

Rapid detection of microorganisms based on active and passive modes of QCM.

作者信息

Farka Zdeněk, Kovář David, Skládal Petr

机构信息

CEITEC MU, Masaryk University, Kamenice 5, 625 00 Brno, Czech Republic.

出版信息

Sensors (Basel). 2014 Dec 23;15(1):79-92. doi: 10.3390/s150100079.

Abstract

Label-free immunosensors are well suited for detection of microorganisms because of their fast response and reasonable sensitivity comparable to infection doses of common pathogens. Active (lever oscillator and frequency counter) and passive (impedance analyzer) modes of quartz crystal microbalance (QCM) were used and compared for rapid detection of three strains of E. coli. Different approaches for antibody immobilization were compared, the immobilization of reduced antibody using Sulfo-SMCC was most effective achieving the limit of detection (LOD) 8 × 104 CFU·mL-1 in 10 min. For the passive mode, software evaluating impedance characteristics in real-time was developed and used. Almost the same results were achieved using both active and passive modes confirming that the sensor properties are not limited by the frequency evaluation method but mainly by affinity of the antibody. Furthermore, reference measurements were done using surface plasmon resonance. Effect of condition of cells on signal was observed showing that cells ruptured by ultrasonication provided slightly higher signal changes than intact microbes.

摘要

无标记免疫传感器因其快速响应以及与常见病原体感染剂量相当的合理灵敏度,非常适合用于微生物检测。采用了石英晶体微天平(QCM)的主动模式(杠杆振荡器和频率计数器)和被动模式(阻抗分析仪)对三株大肠杆菌进行快速检测并比较。比较了不同的抗体固定方法,使用磺基琥珀酰亚胺基4-(N-马来酰亚胺甲基)环己烷-1-羧酸酯(Sulfo-SMCC)固定还原抗体最为有效,在10分钟内实现了8×10⁴CFU·mL⁻¹的检测限(LOD)。对于被动模式,开发并使用了实时评估阻抗特性的软件。主动模式和被动模式获得了几乎相同的结果,证实传感器特性不受频率评估方法限制,主要受抗体亲和力限制。此外,使用表面等离子体共振进行了参考测量。观察到细胞状态对信号的影响,结果表明经超声破碎的细胞比完整微生物产生的信号变化略高。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f1e/4327008/47d10d42867f/sensors-15-00079f1.jpg

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