Tran Vien Thi, Ju Heongkyu
Department of Physics, Gachon University, Seongnam-si, Gyeonggi-do 13120, Korea.
Gachon Bionano Research Institute, Gachon University, Seongnam-si, Gyeonggi-do 13120, Korea.
Biomedicines. 2021 Apr 21;9(5):448. doi: 10.3390/biomedicines9050448.
This work demonstrates the quantitative assay of cardiac Troponin I (cTnI), one of the key biomarkers for acute cardiovascular diseases (the leading cause of death worldwide) using the fluorescence-based sandwich immune reaction. Surface plasmon coupled emission (SPCE) produced by non-radiative coupling of dye molecules with surface plasmons being excitable via the reverse Kretschmann format is exploited for fluorescence-based sandwich immunoassay for quantitative detection of cTnI. The SPCE fluorescence chip utilizes the gold (2 nm)-silver (50 nm) bimetallic thin film, with which molecules of the dye Alexa 488 (conjugated with detection antibodies) make a near field coupling with the plasmonic film for SPCE. The experimental results find that the SPCE greatly improves the sensitivity via enhancing the fluorescence signal (up to 50-fold) while suppressing the photo-bleaching, permitting markedly enhanced signal-to-noise ratio. The limit of detection of 21.2 ag mL (atto-gram mL) is obtained, the lowest ever reported to date amid those achieved by optical technologies such as luminescence and label-free optical sensing techniques. The features discovered such as ultrahigh sensitivity may prompt the presented technologies to be applied for early diagnosis of cTnI in blood, particularly for emergency medical centers overloaded with patients with acute myocardial infarction who would suffer from time-delayed diagnosis due to insufficient assay device sensitivity.
这项工作展示了使用基于荧光的夹心免疫反应对心肌肌钙蛋白I(cTnI)进行定量测定,cTnI是急性心血管疾病(全球主要死因)的关键生物标志物之一。利用通过反向Kretschmann形式可激发的染料分子与表面等离子体的非辐射耦合产生的表面等离子体耦合发射(SPCE),用于基于荧光的夹心免疫测定以定量检测cTnI。SPCE荧光芯片利用金(2纳米)-银(50纳米)双金属薄膜,与检测抗体偶联的染料Alexa 488分子与等离子体薄膜进行近场耦合以实现SPCE。实验结果发现,SPCE通过增强荧光信号(高达50倍)同时抑制光漂白极大地提高了灵敏度,从而显著提高了信噪比。获得了21.2 ag/mL(阿托克/毫升)的检测限,这是迄今为止在诸如发光和无标记光学传感技术等光学技术所达到的检测限中最低的。所发现的超高灵敏度等特性可能促使所提出的技术应用于血液中cTnI的早期诊断,特别是对于因检测设备灵敏度不足而导致急性心肌梗死患者诊断延迟的急诊医疗中心。