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基于分支滚环扩增的用于胃癌外泌体检测的简单荧光适体传感器。

A simple fluorescence aptasensor for gastric cancer exosome detection based on branched rolling circle amplification.

机构信息

State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, National Demonstration Center for Experimental Biomedical Engineering Education (Southeast University), Southeast University, Nanjing 210096, P. R. China.

Economical Forest Cultivation and Utilization of 2011 Collaborative Innovation Center in Hunan Province, Hunan Key Laboratory of Biomedical Nanomaterials and Devices; Hunan University of Technology, Zhuzhou 412007, P. R. China.

出版信息

Nanoscale. 2020 Jan 28;12(4):2445-2451. doi: 10.1039/c9nr08747h. Epub 2020 Jan 2.

Abstract

Exosomes are membrane nanovesicles carrying molecular information that may reflect the biological and genetic characteristics of their parent cells. Numerous studies have demonstrated the potential of exosomes as noninvasive cancer biomarkers. Hence, specific detection of cancer cell-derived exosomes is of significant importance. Here, we developed a fluorescence assay for the determination of gastric cancer exosomes based on branched rolling circle amplification (BRCA) and an aptamer to target specific exosomes. The designed padlock probe was cyclized after incubation with an aptamer binding with the target exosome. BRCA was triggered by adding a second primer and the resulting long tandem double-stranded DNA product was detected using SYBR Green I as the fluorescent dye. This method demonstrated a high specificity for target exosomes with a detection limit of 4.27 × 10 exosomes per mL. Moreover, plasma from gastric cancer patients was tested to verify the clinical applicability of this assay. Our results demonstrated that this aptamer-based biosensor may show potential for the early diagnosis of gastric cancer.

摘要

外泌体是携带分子信息的膜纳米囊泡,可能反映其亲本细胞的生物学和遗传学特征。大量研究表明外泌体作为非侵入性癌症生物标志物的潜力。因此,对癌细胞衍生的外泌体进行特异性检测具有重要意义。在这里,我们开发了一种基于分支滚环扩增(BRCA)和适体的荧光测定法来测定胃癌外泌体。设计的发夹探针在与靶外泌体结合的适体孵育后环化。通过添加第二个引物触发 BRCA,并用 SYBR Green I 作为荧光染料检测所得长串联双链 DNA 产物。该方法对靶外泌体具有高特异性,检测限为每毫升 4.27×10 个外泌体。此外,还检测了胃癌患者的血浆以验证该测定法的临床适用性。我们的结果表明,这种基于适体的生物传感器可能显示出用于早期诊断胃癌的潜力。

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