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基于构象开关诱导杂交的荧光适体传感器,用于简便检测β-淀粉样寡聚物。

Fluorescent aptasensor based on conformational switch-induced hybridization for facile detection of β-amyloid oligomers.

机构信息

School of Pharmacy, College of Pharmacy, Kaohsiung Medical University, Kaohsiung, Taiwan.

Department of Pediatrics, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan.

出版信息

Anal Bioanal Chem. 2022 Nov;414(28):8155-8165. doi: 10.1007/s00216-022-04350-7. Epub 2022 Sep 30.

Abstract

Aβ oligomers (AβO) are a dominant biomarker for early Alzheimer's disease diagnosis. A fluorescent aptasensor coupled with conformational switch-induced hybridization was established to detect AβO. The fluorescent aptasensor is based on the interaction of fluorophore-labeled AβO-specific aptamer (FAM-Apt) against its partly complementary DNA sequence on the surface of magnetic beads (cDNA-MBs). Once the FAM-Apt binds to AβO, the conformational switch of FAM-Apt increases the tendency to be captured by cDNA-MBs. This causes a descending fluorescence of supernatant, which can be utilized to determine the levels of AβO. Thus, the base-pair matching above 12 between FAM-Apt and cDNA-MBs with increasing hybridizing free energies reached the ascending fluorescent signal equilibrium. The optimized aptasensor showed linearity from 1.7 ng mL to 85.1 (R = 0.9977) with good recoveries (79.27-109.17%) in plasma. Furthermore, the established aptasensor possesses rational selectivity in the presence of monomeric Aβ, fibrotic Aβ, and interferences. Therefore, the developed aptasensor is capable of quantifying AβO in human plasma and possesses the potential to apply in clinical cases.

摘要

β 淀粉样蛋白寡聚体 (AβO) 是早期阿尔茨海默病诊断的主要生物标志物。本研究建立了一种荧光适体传感器,该传感器结合构象开关诱导杂交,用于检测 AβO。荧光适体传感器基于荧光标记的 AβO 特异性适体 (FAM-Apt) 与磁珠表面部分互补 DNA 序列 (cDNA-MBs) 的相互作用。一旦 FAM-Apt 与 AβO 结合,FAM-Apt 的构象开关就会增加被 cDNA-MBs 捕获的趋势。这导致上清液荧光下降,可用于确定 AβO 的水平。因此,随着杂交自由能增加,FAM-Apt 与 cDNA-MBs 之间的碱基对匹配超过 12 个,达到上升荧光信号平衡。优化后的适体传感器在血浆中的线性范围为 1.7 ng mL 至 85.1(R = 0.9977),回收率良好(79.27-109.17%)。此外,该建立的适体传感器在存在单体 Aβ、纤维状 Aβ 和干扰物的情况下具有合理的选择性。因此,开发的适体传感器能够定量人血浆中的 AβO,具有应用于临床病例的潜力。

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