Kintzios S, Bem F, Mangana O, Nomikou K, Markoulatos P, Alexandropoulos N, Fasseas C, Arakelyan V, Petrou A-L, Soukouli K, Moschopoulou G, Yialouris C, Simonian A
Laboratory of Plant Physiology, Agricultural University of Athens, Iera Odos 75, 11855, Athens, Greece.
Biosens Bioelectron. 2004 Nov 1;20(4):907-16. doi: 10.1016/j.bios.2004.04.009.
The Bioelectric Recognition Assay (BERA) is a whole-cell based biosensing system that detects the electric response of cultured cells, suspended in a gel matrix, to various ligands, which bind to the cell and/or affect its physiology. Previous studies have demonstrated the potential application of this method for rapid, inexpensive detection of viruses in a crude sample. However, the understanding, so far, of the fundamental processes that take place during cell-virus interactions within the probe has been rather limited. In the present study, we combined electrophysiological and fluorescence microscopical assays, so that we can prove that animal and plant cells immobilized in BERA sensors respond to different viruses primarily by changing their membrane potential. The response of immobilized cells against different viruses did not depend on the virus ability to penetrate the cell, but was modified after binding each virus to a virus-specific antibody or removal of its coat protein after treatment with a protease. Consequently, we were able to assay the presence of a virus in its complete form or fragments thereof. Combination of immunological recognition with the electrophysiological response of immobilized cells allows for a considerable increase of the specificity of the BERA biosensory assay. In addition, rather than simply detect the presence of a protein or genomic sequence, the method can help gain information on the bioactivity of a virus.
生物电识别分析（BERA）是一种基于全细胞的生物传感系统，它能检测悬浮在凝胶基质中的培养细胞对各种配体的电反应，这些配体与细胞结合和/或影响其生理功能。先前的研究已经证明了该方法在快速、廉价检测粗样品中病毒方面的潜在应用。然而，到目前为止，对于探针内细胞与病毒相互作用过程中发生的基本过程的了解还相当有限。在本研究中，我们结合了电生理和荧光显微镜分析，以便能够证明固定在BERA传感器中的动植物细胞主要通过改变其膜电位对不同病毒作出反应。固定细胞对不同病毒的反应并不取决于病毒穿透细胞的能力，而是在每种病毒与病毒特异性抗体结合或用蛋白酶处理后去除其衣壳蛋白后发生改变。因此，我们能够检测完整形式的病毒或其片段的存在。免疫识别与固定细胞的电生理反应相结合，可显著提高BERA生物传感分析的特异性。此外，该方法不仅能简单地检测蛋白质或基因组序列的存在，还能帮助获取有关病毒生物活性的信息。
