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使用截短适配体和SYBR Green I对程序性死亡配体1进行无标记检测以评估预后

Label-Free Detection of Programmed Death-Ligand 1 for Prognosis Using a Truncated Aptamer and SYBR Green I.

作者信息

Li Ling, Li Zheng-Ling, Wang Wei, Long Xin-Xin, Liu Ping, Wang Guo-Tian, Sun Shi-Qing, Du Rong-Lian

机构信息

Department of Oncology, Tengzhou Central People's Hospital, Tengzhou, Shandong, 277599, China.

Tengzhou Maternal and Child Health Hospital, Tengzhou, Shandong, 277599, China.

出版信息

J Fluoresc. 2024 Oct 1. doi: 10.1007/s10895-024-03960-x.

DOI:10.1007/s10895-024-03960-x
PMID:39352679
Abstract

The rapid and accurate detection of programmed death-ligand 1 (PD-L1) expression is of great value in the diagnosis and treatment of tumors. ELISA-based traditional method is the gold standard for protein detection, but there are still some shortcomings, especially the antigen-antibody dependence, greatly increased the detection time and cost. This work constructed a label-free fluorescent probe for rapid and sensitive detection of PD-L1 using a truncated aptamer as recognition molecules and double-stranded DNA specific dyes (SYBR Green I) as signal units. After a series of optimization conditions, this probe has good detection capability for PD-L1 in buffer solution with the detection limit as low as 0.68 ng/mL. Due to the specific recognition ability of aptamer and target, this method also has good selectivity for PD-L1 detection. The recovery of PD-L1 in human serum samples ranges from 86.20 to 96.36%. Compared with other methods, this strategy does not need to be marked, and does not need other complex design and purification process, but simple operation process and strong anti-interference ability. The whole detection process can be completed within 20 min and has good application prospect. This work will provide reference for drug dosage and prognosis evaluation of specific tumor therapy.

摘要

程序性死亡配体1(PD-L1)表达的快速准确检测在肿瘤的诊断和治疗中具有重要价值。基于酶联免疫吸附测定(ELISA)的传统方法是蛋白质检测的金标准,但仍存在一些缺点,尤其是抗原-抗体依赖性,大大增加了检测时间和成本。本研究构建了一种无标记荧光探针,以截短适配体作为识别分子,双链DNA特异性染料(SYBR Green I)作为信号单元,用于快速灵敏地检测PD-L1。经过一系列优化条件后,该探针对缓冲溶液中的PD-L1具有良好的检测能力,检测限低至0.68 ng/mL。由于适配体与靶标的特异性识别能力,该方法对PD-L1检测也具有良好的选择性。人血清样本中PD-L1的回收率为86.20%至96.36%。与其他方法相比,该策略无需标记,也无需其他复杂的设计和纯化过程,操作过程简单且抗干扰能力强。整个检测过程可在20分钟内完成,具有良好的应用前景。本研究将为特定肿瘤治疗的药物剂量和预后评估提供参考。

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