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基于水凝胶的适体传感器通过共振瑞利散射检测凝血酶。

Hydrogel based aptasensor for thrombin sensing by Resonance Rayleigh Scattering.

机构信息

Department of Chemistry, Faculty of Science, Shahid Chamran University of Ahvaz, Ahvaz, Iran.

Department of Chemistry, Faculty of Science, Shahid Chamran University of Ahvaz, Ahvaz, Iran.

出版信息

Anal Chim Acta. 2019 Nov 4;1079:180-191. doi: 10.1016/j.aca.2019.06.049. Epub 2019 Jun 25.

Abstract

In this research, a novel Resonance Rayleigh Scattering (RRS) aptasensor was developed for thrombin monitoring using in-situ synthesized and embedded Au nanoparticles (AuNPs) into poly vinyl alcohol -borax hydrogel (PBH). Thiolated-thrombin binding aptamer (thiolated-TBA) was attached to the surface of AuNPs embedded into PBH to design the PBH-aptasensor for thrombin detection (thiolated-TBA@AuNPs-PBH). To verify the characteristic and morphology of PBH nanocomposite, energy dispersive X-ray analysis, TEM, average particle size analizer and UV-Vis spectra were performed. The difference in RRS intensities in the absence and presence of thrombin was calculated and selected as the monitoring signal. Effect of different parameters on the RRS signal was investigated at excitation wavelength of 500 nm. Under the approved conditions, the linear detection range was validated over the concentration of 0.70 pM- 0.02 μM. The limit of detection based on 3S was 0.10 pM. The relative standard deviation for 5.6 pM and 3.6 nM were 4.0 and 2.7% (n = 10), respectively. The proposed aptasensor was successfully applied as an experimental model for thrombin detection in serum samples of healthy volunteers with acceptable results.

摘要

在这项研究中,开发了一种新型的共振瑞利散射(RRS)适体传感器,用于通过原位合成和嵌入聚 乙烯醇-硼砂水凝胶(PBH)中的金纳米粒子(AuNPs)来监测凝血酶。将巯基化凝血酶结合适体(thiolated-TBA)附着到嵌入 PBH 的 AuNPs 表面,设计用于凝血酶检测的 PBH-适体传感器(thiolated-TBA@AuNPs-PBH)。为了验证 PBH 纳米复合材料的特性和形态,进行了能量色散 X 射线分析、TEM、平均粒径分析器和紫外可见光谱分析。计算了在没有和存在凝血酶的情况下 RRS 强度的差异,并将其选为监测信号。在 500nm 的激发波长下,研究了不同参数对 RRS 信号的影响。在批准的条件下,验证了线性检测范围在 0.70 pM-0.02 μM 的浓度范围内。基于 3S 的检测限为 0.10 pM。对于 5.6 pM 和 3.6 nM,相对标准偏差分别为 4.0%和 2.7%(n=10)。该适体传感器成功应用于健康志愿者血清样品中凝血酶检测的实验模型,结果令人满意。

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