School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou 510006, China.
National and Local United Engineering Lab of Druggability and New Drugs Evaluation, School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou 510006, China.
ACS Sens. 2024 Apr 26;9(4):2083-2090. doi: 10.1021/acssensors.4c00117. Epub 2024 Mar 25.
The early diagnosis of acute myocardial infarction (AMI) is dependent on the combined feedback of multiple cardiac biomarkers. However, it remains challenging to precisely detect multicardiac biomarkers in complex blood early due to the lack of sensitive and specific diagnostic indicators and the low abundance and small size of associated biomarkers with high specificity (such as microRNAs). To make matters worse, spectral overlap significantly limits the multiplex analysis of cardiac biomarkers by fluorescent probes, leading to bias in the diagnosis of myocardial infarction. Herein, we developed a method for simultaneous detection of miRNAs and protein biomarkers using size- and color-coded microbeads that carry signature for target capture. We also constructed a microfluidic chip with different spacer arrays that segregate these microbeads in different chip regions according to their size to produce signature signals, indicating the level of different biomarkers. The signals on the microbeads were hugely amplified by catalytic hairpin assembly and rolling circle amplification. Notably, this strategy enables the simultaneous and in situ sensitive profiling of six kinds of biomarkers via adding two different fluorescent labels, removing the limitations of spectral overlap. We envision that the strategy has great potential for application in clinical diagnosis for AMI.
急性心肌梗死 (AMI) 的早期诊断依赖于多种心脏生物标志物的综合反馈。然而,由于缺乏敏感和特异性的诊断指标,以及与高特异性相关的生物标志物(如 microRNAs)的丰度低、体积小,因此早期在复杂血液中精确检测多心脏生物标志物仍然具有挑战性。更糟糕的是,光谱重叠严重限制了荧光探针对心脏生物标志物的多重分析,导致对心肌梗死的诊断出现偏差。在此,我们开发了一种使用带有目标捕获特征的大小和颜色编码微珠同时检测 miRNA 和蛋白质生物标志物的方法。我们还构建了一种带有不同间隔物阵列的微流控芯片,根据微珠的大小将它们分离到不同的芯片区域中以产生特征信号,从而指示不同生物标志物的水平。微珠上的信号通过催化发夹组装和滚环扩增得到极大放大。值得注意的是,该策略通过添加两种不同的荧光标记物,实现了六种生物标志物的同时和原位敏感分析,消除了光谱重叠的限制。我们设想该策略在 AMI 的临床诊断中有很大的应用潜力。