首页> 外文会议>Italian Conference on Sensors and Microsystems >DEVELOPMENT OF AN APTAMER-BASED ELECTROCHEMICAL SANDWICH ASSAY FOR THE DETECTION OF A CLINICAL BIOMARKER
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DEVELOPMENT OF AN APTAMER-BASED ELECTROCHEMICAL SANDWICH ASSAY FOR THE DETECTION OF A CLINICAL BIOMARKER

机译:用于检测临床生物标志物的适体基电化学夹心测定的研制

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A disposable electrochemical assay involving magnetic particles and carbon-based screen-printed electrodes (SPEs) is developed for the detection of C Reactive Protein (CRP). CRP is a plasma protein and is among the most expressed proteins in acute phase inflammation cases, being a known biomarker for inflammatory states. The assay is based on a sandwich format in which a biotinylated RNA aptamer is immobilised onto streptavidin coated magnetic beads and then coupled to the same biotinylated aptamer. A streptavidin-alkaline phosphatase solution is then added to beads and after the recognition biotin-streptavidin, the modified magnetic beads are captured by a magnet on the surface of a graphite working electrode and the electrochemical detection is thus achieved through the addition of the AP substrate (α-naphthyl-phosphate) and cc-naphthol produced during the enzymatic reaction is detected using Differential Pulse Voltammetry (DPV) The performance of the assay in terms of sensitivity, reproducibility and selectivity are studied in buffer. The detection limit (LOD) is 2.0-10-2 mg/L and the average coefficient of variation (CV) results 9%. The LOD found is comparable with that reported by ELISA and it was much lower than the clinically useful borderline value (8 mg/L).
机译:开发涉及磁性颗粒和基于碳的丝网印刷电极(SPES)的一次性电化学测定以检测C反应蛋白(CRP)。 CRP是血浆蛋白质,是急性期炎症病例中最表达的蛋白质中,是炎症状态的已知生物标志物。该测定是基于其中生物素化的RNA适体固定在链亲和素包被的磁珠,然后耦合到相同的生物素化的适配体夹心形式。然后将链霉抗生物素蛋白 - 碱性磷酸酶溶液加入珠粒中,在识别生物素 - 链霉抗生物素蛋白之后,通过添加AP衬底,通过添加AP衬底来捕获修饰的磁珠。通过添加AP衬底来实现电化学检测使用差分脉冲伏安法(DPV)检测在酶反应期间产生的α-萘基 - 磷酸酯)和CC-萘酚,在缓冲液中研究了测定的性能,在缓冲液中研究了再现性和选择性。检测限(LOD)为2.0-10-2 mg / L,平均变异系数(CV)结果为9%。发现的LOD与ELISA报道的床单可比,但远低于临床有用的边界值(8mg / L)。

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