State Key Laboratory Base of Eco-Chemical Engineering, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao, P. R. China.
Clin Chem. 2012 Feb;58(2):384-90. doi: 10.1373/clinchem.2011.173195. Epub 2011 Dec 15.
High sensitivity of analysis is constantly in demand in biomedical research and clinical diagnosis. In recent years aptamer-based analytical methods have been developed for protein detection. We developed a cascade signal amplification strategy based on molecular switches and aptamers to improve protein detection.
Our cascade signal amplification strategy based on molecular switches and aptamers consisted of 2 steps, including the recognition and the triggering of a polymerase reaction. The procedure was designed to simplify the analysis by detecting trace amounts of target isothermally, in real time, and in a homogeneous solution. We applied this method to measure thrombin in human serum samples.
This cascade signal amplification strategy exhibited a linear response in thrombin concentration from 0.3 to 10 nmol/L, with a detection limit of 1.7 × 10(-10) mol/L within 60 min. Results of the analysis of thrombin in human serum diluted 1:1 appeared to be linear, as was observed in buffer, in the tested concentration range of 0.3-10 nmol/L.
The aptameric sensor provides promising potential for detecting and screening trace concentrations of biomarkers in complex matrices for clinical applications.
在生物医学研究和临床诊断中，对分析的高灵敏度的需求不断增加。近年来，基于适体的分析方法已经被开发出来用于蛋白质检测。我们开发了一种基于分子开关和适体的级联信号放大策略，以提高蛋白质检测的灵敏度。
我们基于分子开关和适体的级联信号放大策略包括 2 个步骤，包括识别和引发聚合酶反应。该程序旨在通过在等温、实时和均相溶液中检测痕量的靶标来简化分析。我们将这种方法应用于测量人血清样品中的凝血酶。
这种级联信号放大策略在凝血酶浓度为 0.3 到 10 nmol/L 之间呈现线性响应，在 60 分钟内检测限为 1.7×10(-10)mol/L。在缓冲液中，在 0.3-10 nmol/L 的测试浓度范围内，人血清中凝血酶的稀释 1:1 的分析结果呈线性。
适体传感器为在复杂基质中检测和筛选痕量生物标志物提供了有前景的潜力，可用于临床应用。
