One-Step Digital Immunoassay for Rapid and Sensitive Detection of Cardiac Troponin I.,ACS Sensors

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One-Step Digital Immunoassay for Rapid and Sensitive Detection of Cardiac Troponin I.
ACS Sensors ( IF 8.2 ) Pub Date : 2020-03-17 , DOI: 10.1021/acssensors.0c00064
Yi Wang ₁ , Yunze Yang ₂ , Chao Chen ₂ , Shaopeng Wang ₂ , Hui Wang ₁ , Wenwen Jing ₂ , Nongjian Tao _{1,

2}

Affiliation

A rapid and sensitive method to detect cardiac troponin I (cTnI) in human blood is critical to the diagnosis and treatment of acute myocardial infarction (AMI). Here, we describe a simple one-step digital immunoassay for single-molecule detection without washing steps. A sample containing cTnI mixed with detection antibody-conjugated gold nanoparticles (AuNPs) is added to a capture antibody-coated sensor surface and the formation of the antibody-cTnI-antibody sandwich is detected by digitally counting the binding of the individual gold nanoparticles to the sensor surface in real time using a bright-field optical imaging setup together with a differential imaging algorithm. The digital immunoassay detects cTnI in undiluted human plasma, which achieves a detection limit of 5.7 ng/L within a detection time of only 10 min, which meets the requirement of current clinical high-sensitivity troponin assay (∼70 ng/L cutoff). We anticipate that the one-step and real-time digital immunoassay can be applied to the detection of other disease biomarkers in blood.

中文翻译：

快速，灵敏地检测心肌肌钙蛋白I的一步式数字免疫分析。

快速，灵敏的检测人血中心肌肌钙蛋白I（cTnI）的方法对于急性心肌梗死（AMI）的诊断和治疗至关重要。在这里，我们描述了一种无需清洗步骤即可进行单分子检测的简单的一步式数字免疫测定方法。将包含cTnI与检测抗体结合的金纳米颗粒（AuNPs）混合的样品添加到捕获抗体包被的传感器表面，并通过数字计算单个金纳米颗粒与单抗的结合来检测抗体-cTnI-抗体三明治的形成。使用明场光学成像装置和差分成像算法实时地对传感器表面进行成像。数字免疫测定法可在未稀释的人类血浆中检测cTnI，在仅10分钟的检测时间内即可达到5.7 ng / L的检测限，可以满足当前临床高灵敏度肌钙蛋白测定（约70 ng / L临界值）的要求。我们预计，一步和实时数字免疫测定法可应用于血液中其他疾病生物标记物的检测。

更新日期：2020-03-17

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