State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province, Hunan University, Changsha 410082, China.
State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province, Hunan University, Changsha 410082, China.
Biosens Bioelectron. 2015 Feb 15;64:161-4. doi: 10.1016/j.bios.2014.08.079. Epub 2014 Sep 3.
Myoglobin (Myo), which is one of the early markers to increase after acute myocardial infarction (AMI), plays a major role in urgent diagnosis of cardiovascular diseases. Hence, monitoring of Myo in point-of-care is fundamental. Here, a novel assay for sensitive and selective detection of Myo was introduced using a personal glucose meter (PGM) as readout. In the presence of Myo, the anti-Myo antibody immobilized on the surface of polystyrene microplate could capture the target Myo. Then the selected aptamer against Myo, which was obtained using our screening process, was conjugated with invertase, and such aptamer-invertase conjugates bound to the immobilized Myo due to the Myo/aptamer interaction. Subsequently, the resulting "antibody-Myo-aptamer sandwich" complex containing invertase conjugates hydrolyzed sucrose into glucose, thus establishing direct correlation between the Myo concentration and the amount of glucose measured by PGM. By employing the enzyme amplification, as low as 50 pM Myo could be detected. This assay also showed high selectivity for Myo and was successfully used for Myo detection in serum samples. This work may provide a simple but reliable tool for early diagnosis of AMI in the world, especially in developing countries.
肌红蛋白(Myo)是急性心肌梗死(AMI)后最早升高的标志物之一,在心血管疾病的紧急诊断中发挥着重要作用。因此,即时检测(Point-of-care testing,POCT)中对 Myo 的监测至关重要。本研究使用个人血糖仪(PGM)作为读出设备,介绍了一种用于检测 Myo 的高灵敏度和高选择性的新方法。在 Myo 存在的情况下,固定在聚苯乙烯微孔板表面的抗 Myo 抗体可以捕获目标 Myo。然后,选择与我们的筛选过程获得的针对 Myo 的适体与蔗糖酶结合,由于 Myo/适体的相互作用,适体-蔗糖酶结合物与固定化 Myo 结合。随后,包含蔗糖酶结合物的“抗体-Myo-适体夹心”复合物将蔗糖水解成葡萄糖,从而建立了 Myo 浓度与 PGM 测量的葡萄糖量之间的直接相关性。通过酶放大作用,可以检测到低至 50 pM 的 Myo。该检测方法对 Myo 具有很高的选择性,并成功用于血清样品中的 Myo 检测。这项工作可能为世界范围内,特别是发展中国家的 AMI 的早期诊断提供一种简单但可靠的工具。