Time-resolved fluorescence resonance energy transfer-based lateral flow immunoassay using a raspberry-type europium particle and a single membrane for the detection of cardiac troponin I
Time-resolved fluorescence resonance energy transfer-based lateral flow immunoassay using a raspberry-type europium particle and a single membrane for the detection of cardiac troponin I
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DOI：
10.1016/j.bios.2020.112284
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发表时间：
2020-09-01
影响因子：
12.6
通讯作者：
Yoon, Hyun C.
中科院分区：
文献类型：
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作者：
Lee, Kyung Won;Kim, Ka Ram;Yoon, Hyun C.
在此，我们报告了一种新的侧流免疫分析（LFIA）系统检测血清中的心肌肌钙蛋白I（cTnI），使用时间分辨荧光共振能量转移（TR-FRET）技术和融合5膜。Fusion 5膜用作LFIA的条带，并且其在没有额外基质（例如样品或缀合垫）的情况下构建。虽然这种构建LFIA试纸条的策略非常简单且具有成本效益，但LFIA仍然不适合分析需要高灵敏度的生物标志物，如cTnI。因此，高灵敏度的TR-FRET技术与基于融合5膜的LFIA条集成。为了实现这一点，覆盖有含铕螯合物的二氧化硅纳米颗粒的微粒被合成为树莓型颗粒并用作荧光供体。金纳米棒（GNR）用作荧光受体颗粒。在基于TR-FRET的LFIA系统中，应进行竞争性免疫测定以满足FRET现象发生所需的条件。因此，荧光信号与cTnI浓度成正比，确保通过测量树莓型铕颗粒与GNR之间的荧光信号可以实现cTnI的定量分析。使用开发的TR-FRET为基础的LFIA系统，cTnI的灵敏检测成功地实现了97 pg/mL的检测限在人血清中。此外，由于可以使用一种基质（fusion 5膜）获得结果，因此开发的LFIA系统可以通过简单的制造过程用于cTnI诊断。
Herein, we report a novel lateral flow immunoassay (LFIA) system for detecting cardiac troponin I (cTnI) in serum using the time-resolved fluorescence resonance energy transfer (TR-FRET) technique and the fusion 5 membrane. The fusion 5 membrane is used as a strip for LFIA, and it is constructed without additional matrices (such as a sample or conjugation pad). Although this strategy for constructing the LFIA strip is quite simple and cost-effective, LFIA is still not suitable for the analysis of biomarkers that require high sensitivity, such as cTnI. Therefore, the highly sensitive TR-FRET technique is integrated with a fusion 5 membrane-based LFIA strip. To accomplish this, a microparticle covered with europium chelate-contained silica nanoparticles is synthesized as a raspberry-type particle and used as a fluorescence donor. A gold nanorod (GNR) is used as a fluorescence acceptor particle. In the TR-FRET-based LFIA system, the competitive immunoassay should be performed to satisfy the condition required for the FRET phenomenon to occur. Therefore, the fluorescence signal is proportional to the cTnI concentration, ensuring a quantitative analysis of cTnI can be accomplished by measuring the fluorescence signal between the raspberry-type europium particles and GNR. Using the developed TR-FRETbased LFIA system, sensitive detection of cTnI is successfully achieved with a limit of detection of 97 pg/mL in human serum. Moreover, because the result can be obtained using one matrix (the fusion 5 membrane), the developed LFIA system can be employed in cTnI diagnosis with a simple manufacturing process.