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基于表面增强拉曼光谱的磁性免疫分析法,利用核壳纳米标签同时检测肌钙蛋白I和心脏型脂肪酸结合蛋白

SERS-based magnetic immunoassay for simultaneous detection of cTnI and H-FABP using core-shell nanotags.

作者信息

Hu Cunming, Ma Li, Guan Ming, Mi Fang, Peng Fei, Guo Chang, Sun Shijiao, Wang Xiaomei, Liu Tingwei, Li Jiutong

机构信息

College of Chemistry and Chemical Engineering, Xinjiang Normal University, Urumqi 830054, China.

College of Pharmacy, Xinjiang Medical University, Urumqi 830011, China.

出版信息

Anal Methods. 2020 Dec 7;12(45):5442-5449. doi: 10.1039/d0ay01564d. Epub 2020 Nov 9.

Abstract

Acute myocardial infarction (AMI) is the single leading cause of worldwide mortality and morbidity. Heart-type fatty acid-binding protein (H-FABP) and cardiac troponin I (cTnI), as biomarkers emerging at different stages of AMI, have complementary advantages in terms of specificity and sensitivity. Therefore, we developed a magnetic immunoassay method based on surface-enhanced Raman scattering (SERS) to detect H-FABP and cTnI simultaneously. Herein, two mutually independent Raman reporter molecules were embedded between a gold core and silver shell and then combined with a tracer antibody to form a SERS immunoprobe. During detection, the SERS immunoprobe, target antigen and capture probe undergo an immune reaction to form a sandwich structure, and then the immune complex was enriched by a specific reaction of streptavidin on the surface of magnetic beads with biotin. Finally, the concentration of biomarkers was quantified by detecting the characteristic Raman peak intensities of the two Raman reporter molecules. Under the optimized conditions, the minimum detection limits of H-FABP and cTnI were 0.6396 and 0.0044 ng mL, respectively. Besides, the target antigen does not cross-react with non-specific proteins, showing good specificity. Therefore, our proposed SERS-based magnetic immunoassay method has the advantages of accuracy, rapidity and good selectivity, and has great potential for early diagnosis of acute myocardial infarction disease.

摘要

急性心肌梗死(AMI)是全球范围内导致死亡和发病的首要单一原因。心脏型脂肪酸结合蛋白(H-FABP)和心肌肌钙蛋白I(cTnI)作为在AMI不同阶段出现的生物标志物,在特异性和敏感性方面具有互补优势。因此,我们开发了一种基于表面增强拉曼散射(SERS)的磁免疫分析方法,用于同时检测H-FABP和cTnI。在此,将两个相互独立的拉曼报告分子嵌入金核和银壳之间,然后与示踪抗体结合形成SERS免疫探针。在检测过程中,SERS免疫探针、靶抗原和捕获探针发生免疫反应形成夹心结构,然后通过磁珠表面的链霉亲和素与生物素的特异性反应富集免疫复合物。最后,通过检测两个拉曼报告分子的特征拉曼峰强度来定量生物标志物的浓度。在优化条件下,H-FABP和cTnI的最低检测限分别为0.6396和0.0044 ng/mL。此外,靶抗原与非特异性蛋白质不发生交叉反应,显示出良好的特异性。因此,我们提出的基于SERS的磁免疫分析方法具有准确性、快速性和良好的选择性等优点,在急性心肌梗死疾病的早期诊断方面具有巨大潜力。

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