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评估全细胞生物传感器分析遗传毒性的计时安培信号检测。

Evaluation of chrono-amperometric signal detection for the analysis of genotoxicity by a whole cell biosensor.

机构信息

Division of Qualitative Hydrology, German Federal Institute of Hydrology (BfG), Am Mainzer Tor 1, Koblenz 56068, Germany.

出版信息

Anal Chim Acta. 2010 Feb 5;659(1-2):122-8. doi: 10.1016/j.aca.2009.11.027. Epub 2009 Nov 18.

Abstract

Electrochemical signal detection can be readily integrated in biosensors and is thus an attractive alternative to optical detection methods. In the field of environmental chemistry and ecotoxicology there is a growing demand for lab-independent devices based on whole cell biosensors for the detection of genotoxic compounds. Because of the broad occurrence of pre-genotoxic compounds that need to be bio-activated, the integration of a system for metabolic activation into such a biosensor is important. The present study evaluates a chrono-amperometric detection method in which para-aminophenyl beta-D-galactopyranoside is used as substrate for a reporter gene assay based on the bacterial SOS-response in comparison to a test system for the determination of genotoxicity in water that is standardized according to the International Organization for Standardization (ISO). The evaluation was done in order to analyze the potential of the electrochemical signal detection to be used as a complementary method for the standard test system and thus to evaluate the usability of electrochemical biosensors for the assessment of genotoxicity of environmental samples. In the present study it is shown that the chrono-amperometric detection of para-aminophenol is specific even in the presence of electro-active species generated by the enzymatic system used for the external bio-activation of contaminants. Under optimized conditions electrochemistry is sufficiently sensitive with a limit of detection that is comparable to the respective ISO-standard.

摘要

电化学信号检测可方便地整合到生物传感器中,因此是光学检测方法的一种有吸引力的替代方法。在环境化学和生态毒理学领域,人们对基于全细胞生物传感器的独立于实验室的设备的需求不断增长,这些设备用于检测遗传毒性化合物。由于需要生物激活的前遗传毒性化合物的广泛存在,因此将代谢激活系统集成到这种生物传感器中非常重要。本研究评估了一种计时电流检测方法,其中对氨基苯-β-D-半乳糖苷被用作基于细菌 SOS 反应的报告基因测定的底物,与根据国际标准化组织 (ISO) 标准化的用于测定水中遗传毒性的测试系统进行比较。进行评估是为了分析电化学信号检测作为标准测试系统的补充方法的潜力,从而评估电化学生物传感器用于评估环境样品遗传毒性的可用性。在本研究中,即使在用于外部生物激活污染物的酶系统产生的电活性物质存在的情况下,对氨基苯酚的计时电流检测也具有特异性。在优化条件下,电化学具有足够的灵敏度,检测限与相应的 ISO 标准相当。

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