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Preliminary study of a low-cost point-of-care testing system using screen-printed biosensors: For early biomarkers detection related to Alzheimer Disease

机译:使用丝网印刷生物传感器的低成本即时检验系统的初步研究:用于与阿尔茨海默氏病相关的早期生物标记检测

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Among neurodegenerative diseases, Alzheimer Disease (AD) represents one of the most widespread pathologies, for which an early diagnosis is still missing. A peculiar expression of an altered conformational isoform of p53 protein was reported to be a potential biomarker able to distinguish AD subjects from healthy population, quantifiable using a blood-based enzyme-linked immunosorbent assay (ELISA). In order to overcome ELISA limitations, related to reliability and to improve sensitivity, this study aimed to realize a low cost highly sensitive portable point-of-care (PoC) testing system based on screen printed electrochemical sensors (SPES). The development of the platform specifically included both the design of the sensing probe and of the electronic circuit devoted to the conditioning and acquisition of the transduced electric signal. In particular, silver, carbon and silver-silver chloride were selected respectively to realize conductive tracks, working and counter electrodes, reference electrode in a three-electrodes configuration focusing on Anodic Stripping Voltammetry (ASV). The conditioning circuit was designed following the scheme for a common potentiostat, and produced as a Printed Circuit Board (PCB). Initial testing of the circuit were performed recording changes in the conductivity of NaCl solution and quantifying electrodes coating with antibodies using Electrochemical Impedance Spectroscopy (EIS) principle. Preliminary results obtained with saline solution, showed the ability of the circuit to give the best response corresponding to low changes in NaCl concentration (sensitivity 0.2 mA/(mg/ml)), suggesting a good sensitivity of the platform. Results from EIS showed the ability of the circuit to discriminate between different concentrations of antibodies coatings (sensitivity 80 μA/μg). The study is on-going and after a proper calibration, the circuit is intended to be optimized to quantify unknown concentration of unfolded p53 in samples of peripheral blood of AD patients, compared results with the one from ELISA analysis, aiming to realize a low cost, easy to use and highly precise platform.
机译:在神经退行性疾病中,阿尔茨海默病(AD)代表最广泛的病理之一,至今仍缺乏早期诊断。据报道,p53蛋白构象同工异型的表达是一种潜在的生物标志物,能够将AD受试者与健康人群区分开,可以使用基于血液的酶联免疫吸附测定(ELISA)进行定量。为了克服与可靠性相关的ELISA局限性并提高灵敏度,本研究旨在实现一种基于丝网印刷电化学传感器(SPES)的低成本高灵敏度便携式即时护理(PoC)测试系统。该平台的开发特别包括传感探头的设计和专门用于调节和获取所转换的电信号的电子电路的设计。特别是,分别选择了银,碳和氯化银-银,以阳极阳极伏安法(ASV)为三电极结构来实现导电走线,工作电极和对电极以及参比电极。调节电路是按照通用恒电位仪的方案设计的,并制成印刷电路板(PCB)。进行电路的初始测试,记录NaCl溶液的电导率变化,并使用电化学阻抗谱(EIS)原理量化涂有抗体的电极。用盐溶液获得的初步结果表明,该电路具有与NaCl浓度低变化(灵敏度0.2 mA /(mg / ml))相对应的最佳响应能力,表明该平台具有良好的灵敏度。 EIS的结果表明,该电路能够区分不同浓度的抗体涂层(灵敏度为80μA/μg)。这项研究正在进行中,经过适当的校准后,可以优化该电路以定量AD患者外周血样品中未折叠的p53浓度,并将其与ELISA分析的结果进行比较,以实现低成本,易于使用且高度精确的平台。

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