骨架和球形核酸协同增强电化学发光纳米传感器基于循环 MicroRNA 水平快速诊断急性心肌梗死,Analytical Chemistry

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骨架和球形核酸协同增强电化学发光纳米传感器基于循环 MicroRNA 水平快速诊断急性心肌梗死
Analytical Chemistry ( IF 6.7 ) Pub Date : 2022-10-05 , DOI: 10.1021/acs.analchem.2c03144
Lin Shi ₁ , Chengbin Liu ₁ , Han Wang ₁ , Jiao Zheng ₁ , Qiwei Wang ₁ , Lili Shi ₂ , Tao Li ₁

Affiliation

循环血液中与急性心肌梗死 (AMI) 相关的 microRNA (miRNA) 已被证明是 AMI 诊断的有前途的生物标志物。检测这些超低水平的 miRNA 通常需要核酸扩增策略来提高灵敏度，但代价是时间成本。鉴于急性心肌梗死发作后的第一个小时是挽救急性心肌梗死患者生命的黄金时间，缩短超灵敏miRNA分析时间对临床急性心肌梗死诊断具有重要意义。为了实现这一目标，我们在这里提出了一种用于快速和超灵敏 miRNA 检测的直接电化学发光 (ECL) 传感策略，避免了耗时的信号放大步骤。目标 miRNA 与两条探针链直接杂交，这两条探针链分别连接到电极上的共价血红素修饰的球形核酸酶 (SNAzyme) 和截短的三角锥体 DNA 纳米平台上。两者均提高了ECL信号，同时降低了背景，从而显着提高了目标miRNA的检测灵敏度。它能够使用 SNAzyme 催化的鲁米诺 H 在 30 分钟内快速检测人血清中上午 10 点的 AMI 相关 miRNA (miR-499)₂ O ₂ ECL反应。然后将该传感策略用于 AMI 诊断，通过以内源性 miR-16 作为内在参考探测患者循环血液中的内源性 miR-499，显示患者和健康人的 miR-499 水平之间存在显着差异（ P < 0.001） .

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Framework and Spherical Nucleic Acids Synergistically Enhanced Electrochemiluminescence Nanosensors for Rapidly Diagnosing Acute Myocardial Infarction Based on Circulating MicroRNA Levels

Acute myocardial infarction (AMI)-related microRNAs (miRNAs) in circulating blood have been proved as promising biomarkers for AMI diagnosis. The detection of these miRNAs at ultralow levels usually requires nucleic acid amplification strategies to improve the sensitivity at the cost of time. Given that the first hour after an AMI attack is the golden time for saving AMI patients’ lives, shortening the time of ultrasensitive miRNA analysis is of great significance for clinical AMI diagnosis. Toward this goal, here we present a direct electrochemiluminescence (ECL) sensing strategy for fast and ultrasensitive miRNA detection, circumventing the time-consuming signal amplification steps. Target miRNAs are directly hybridized with two probe strands that are attached to a covalently hemin-modified spherical nucleic acid enzyme (SNAzyme) and a truncated triangular pyramid DNA nanoplatform on the electrode, respectively. Both of them improve the ECL signal and meanwhile reduce the background, thereby remarkably promoting the detection sensitivity of target miRNAs. It enables the rapid detection of an AMI-related miRNA (miR-499) at 10 aM in human serum within 30 min using the SNAzyme-catalyzed luminol-H₂O₂ ECL reaction. This sensing strategy is then utilized for AMI diagnosis via probing endogenous miR-499 in patients’ circulating blood with endogenous miR-16 as an intrinsic reference, showing a significant difference (P < 0.001) between the miR-499 levels of patients and the healthy.

更新日期：2022-10-05

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