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用于特异性检测胰腺癌外泌体miRNA10b的集成场效应晶体管传感微系统

Integrated FET sensing microsystem for specific detection of pancreatic cancer exosomal miRNA10b.

作者信息

Yu Yi, Liang Chunzi, Wan Qiang-Qiang, Jin Dan, Liu Xi, Zhang Zhiyong, Sun Zhong-Yue, Zhang Guo-Jun

机构信息

School of Laboratory Medicine, Hubei University of Chinese Medicine, 16 Huangjia Lake West Road, Wuhan, 430065, China.

Wuhan First Hospital, Wuhan, 430022, China.

出版信息

Anal Chim Acta. 2023 Dec 15;1284:341995. doi: 10.1016/j.aca.2023.341995. Epub 2023 Nov 2.

Abstract

Tumor-derived exosome (TD-Ex) serves as a crucial early diagnostic biomarker of pancreatic cancer (PC). However, accurate identification of TD-Ex from PC is still a challenging work. In this paper, a detection microsystem that integrates magnetic separation and FET biosensor is developed, which is capable of selectively separating TD-Ex of PC from the plasma and detecting exosomal miRNA10b in a sensitive and specific manner. The magnetic beads were functionalized with dual antibody (GPC-1 antibody and EpCAM antibody), enabling selective recognition and capture of PC-derived exosomes. On the other hand, a peptide nucleic acid (PNA)- functionalized reduced graphene oxide field-effect transistor (RGO FET) biosensor was subsequently utilized to detect the exosomal miRNA10b, which is highly expressed in PC- derived exosomes. This system could achieve a low detection limit down to 78 fM, and selectively identify miRNA10b from single-base mismatched miRNA. In addition, 40 clinical plasma samples were tested with this microsystem, and the results indicate that it could effectively distinguish PC patients from healthy individuals. The assay combines specific capture and enrichment of PC-derived exosomes with sensitive and selective detection of exosomal miRNA, showing its potential to be used as an effective scheme for PC early diagnosis.

摘要

肿瘤衍生外泌体(TD-Ex)是胰腺癌(PC)的一种关键早期诊断生物标志物。然而,从胰腺癌中准确识别TD-Ex仍然是一项具有挑战性的工作。本文开发了一种集成磁分离和场效应晶体管(FET)生物传感器的检测微系统,该系统能够从血浆中选择性分离胰腺癌的TD-Ex,并以灵敏且特异的方式检测外泌体miRNA10b。用双抗体(GPC-1抗体和EpCAM抗体)对磁珠进行功能化处理,从而实现对胰腺癌衍生外泌体的选择性识别和捕获。另一方面,随后利用肽核酸(PNA)功能化的还原氧化石墨烯场效应晶体管(RGO FET)生物传感器检测在胰腺癌衍生外泌体中高表达的外泌体miRNA10b。该系统能够实现低至78飞摩尔(fM)的检测限,并能从单碱基错配的miRNA中选择性识别miRNA10b。此外,用该微系统对40份临床血浆样本进行了检测,结果表明它能够有效区分胰腺癌患者和健康个体。该检测方法将胰腺癌衍生外泌体的特异性捕获和富集与外泌体miRNA的灵敏且选择性检测相结合,显示出其作为胰腺癌早期诊断有效方案的潜力。

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