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基于 SERS 磁免疫分析的白细胞介素 6 的快速、高灵敏和定量检测。

Rapid, highly sensitive and quantitative detection of interleukin 6 based on SERS magnetic immunoassay.

机构信息

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

Shanghai Simp Bio-science Co., Ltd, 201318, China.

出版信息

Anal Methods. 2021 Apr 22;13(15):1823-1831. doi: 10.1039/d0ay02304c.

Abstract

Sepsis is a systemic inflammatory response syndrome caused by infection, and is a common disease in intensive care units (ICUs), whereby the mortality rate is as high as 30% to 50%. Early diagnosis and treatment can significantly reduce the mortality rate of patients with sepsis. We have developed a method based on SERS for the rapid and quantitative detection of IL-6. Using the principle of double antibody sandwich, the core-shell nanoparticles embedded with a Raman reporter (Au@4MBA@Ag NPs) are coupled to the tracer antibody, while the biotin was coupled to the capture antibody to form an antibody-antigen-antibody sandwich structure with the antigen during detection of the structure. Streptavidin (SA) and biotin had a strong affinity, and the sandwich structure was captured by SA magnetic beads and detected by Raman spectroscopy under the enrichment of an external magnetic field. The results showed a good linear relationship between the Raman signal and the concentration of IL-6 in the concentration range of 0-1000 pg mL-1 (r = 0.9997) with a limit of detection of 1.6 pg mL-1. Also, the recovery rate of standard addition was 93.9-99.1%, and the coefficient of variation intra-assay and inter-assay of the three batches of reagents was less than 15%. Furthermore, it showed excellent specificity with procalcitonin (PCT, 20 ng mL-1) and C-reactive protein (CRP, 100 μg mL-1) and had no cross-reactivity. Except for bilirubin (2 mg mL-1) and hemoglobin (10 mg mL-1), other common interferences in the serum did not interfere, showing good anti-interference performance. Moreover, 57 clinical serum samples were detected via the chemiluminescence method simultaneously, and the detection results showed a good correlation (R2 = 0.9793, P < 0.01). There was no significant difference between the two performances (P > 0.05). The proposed method has numerous advantages such as high sensitivity, wide linear range, short detection time and simple operation, which provide a new technical reference for the clinical detection of sepsis biomarkers.

摘要

脓毒症是一种由感染引起的全身性炎症反应综合征,是重症监护病房(ICU)中的常见疾病,其死亡率高达 30%至 50%。早期诊断和治疗可以显著降低脓毒症患者的死亡率。我们开发了一种基于 SERS 的快速定量检测 IL-6 的方法。利用双抗体夹心原理,将嵌入拉曼报告分子(Au@4MBA@Ag NPs)的核壳纳米粒子与示踪抗体偶联,而生物素与捕获抗体偶联,在检测结构时形成抗原-抗体-抗原夹心结构。链霉亲和素(SA)与生物素有很强的亲和力,在外部磁场的富集作用下,夹心结构被 SA 磁珠捕获,并通过拉曼光谱检测。结果表明,在 0-1000 pg mL-1 的浓度范围内,拉曼信号与 IL-6 浓度呈良好的线性关系(r = 0.9997),检出限为 1.6 pg mL-1。此外,标准加入的回收率为 93.9%-99.1%,三批试剂的日内和日间变异系数均小于 15%。此外,它与降钙素原(PCT,20 ng mL-1)和 C 反应蛋白(CRP,100 μg mL-1)表现出优异的特异性,且无交叉反应。除胆红素(2 mg mL-1)和血红蛋白(10 mg mL-1)外,血清中其他常见干扰物质不干扰,表现出良好的抗干扰性能。同时,采用化学发光法检测了 57 例临床血清样本,检测结果相关性良好(R2 = 0.9793,P < 0.01)。两种性能之间无显著差异(P > 0.05)。该方法具有灵敏度高、线性范围宽、检测时间短、操作简单等优点,为脓毒症生物标志物的临床检测提供了新的技术参考。

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