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基于聚集诱导发射纳米粒子的数字微流控芯片工作站快速灵敏检测人心型脂肪酸结合蛋白。

Rapid and sensitive detection of heart-type fatty acid binding protein using aggregation-induced emission nanoparticles on digital microfluidics workstation.

机构信息

Department of Laboratory Medicine, Guangzhou First People's Hospital, the Second Affiliated Hospital, School of medicine, South China University of Technology, Guangzhou, 510180, China.

Institute of Antibody Engineering, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, 510515, China.

出版信息

Biosens Bioelectron. 2024 Oct 15;262:116563. doi: 10.1016/j.bios.2024.116563. Epub 2024 Jul 10.

Abstract

Early and rapid diagnostic of acute myocardial infarction (AMI) during its developing stage is crucial due to its high fatality rate. Heart-type fatty acid binding protein (h-FABP) is an ideal biomarker for the quantitative diagnosis of AMI, surpassing traditional markers such as myoglobin, creatine phosphokinase-MB, and troponin in terms of sensitivity, specificity, and prognostic value. To obtain diagnostic and prognostic information, a precise and fully quantitative measurement of h-FABP is essential, typically achieved through an immunosorbent assay like the enzyme-linked immunosorbent assay. Nevertheless, this method has several limitations, including extended detection time, complex assay procedures, the necessity for skilled technicians, and challenges in implementing automated detection. This research introduces a novel biosensor, utilizing aggregation-induced emission nanoparticles (AIENPs) and integrated with a digital microfluidic (DMF) workstation, designed for the sensitive, rapid, and automated detection of h-FABP in low-volume serum samples. AIENPs and magnetic beads in nanoscale were served as the capture particles and the fluorescent probe, which were linked covalently to anti-h-FABP antibodies respectively. The approach was based on a sandwich immunoassay and performed on a fully automated DMF workstation with assay time by 15 min. We demonstrated the determination of h-FABP in serum samples with detection limit of 0.14 ng/mL using this biosensor under optimal condition. Furthermore, excellent correlations (R = 0.9536, n = 50) were obtained between utilizing this biosensor and commercialized ELISA kits in clinical serum detecting. These results demonstrate that our flexible and reliable biosensor is suitable for direct integration into clinical diagnostics, and it is expected to be promising diagnostic tool for early detection and screening tests as well as prognosis evaluation for AMI patients.

摘要

早期和快速诊断急性心肌梗死(AMI)在其发展阶段是至关重要的,因为它的死亡率很高。心脏型脂肪酸结合蛋白(h-FABP)是 AMI 定量诊断的理想生物标志物,在灵敏度、特异性和预后价值方面超过了肌红蛋白、肌酸磷酸激酶-MB 和肌钙蛋白等传统标志物。为了获得诊断和预后信息,需要精确和完全定量测量 h-FABP,通常通过酶联免疫吸附测定等免疫吸附测定来实现。然而,这种方法存在几个局限性,包括检测时间延长、检测程序复杂、需要熟练的技术人员以及在实现自动化检测方面的挑战。本研究引入了一种新型生物传感器,利用聚集诱导发射纳米粒子(AIENPs)并与数字微流控(DMF)工作站集成,用于在小体积血清样本中灵敏、快速和自动检测 h-FABP。纳米级的 AIENPs 和磁性珠被用作捕获颗粒和荧光探针,分别与抗 h-FABP 抗体共价连接。该方法基于夹心免疫测定,并在全自动 DMF 工作站上进行,测定时间为 15 分钟。在最佳条件下,我们使用该生物传感器在血清样本中测定 h-FABP 的检测限为 0.14ng/mL。此外,我们在临床血清检测中利用该生物传感器和商业化 ELISA 试剂盒获得了极好的相关性(R=0.9536,n=50)。这些结果表明,我们的灵活可靠的生物传感器非常适合直接集成到临床诊断中,有望成为 AMI 患者早期检测、筛查试验和预后评估的有前途的诊断工具。

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