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用于细胞外囊泡微小RNA的高效自动化分离技术

Highly efficient and automated isolation technology for extracellular vesicles microRNA.

作者信息

Di Kaili, Fan Boyue, Gu Xinrui, Huang Rongrong, Khan Adeel, Liu Chang, Shen Han, Li Zhiyang

机构信息

Department of Laboratory Medicine, Affiliated Drum Tower Hospital, Medical School of Nanjing University, Nanjing, China.

Jiangsu Key Laboratory of Medical Science and Laboratory Medicine, School of Medicine, Jiangsu University, Zhenjiang, China.

出版信息

Front Bioeng Biotechnol. 2022 Aug 10;10:948757. doi: 10.3389/fbioe.2022.948757. eCollection 2022.

Abstract

MicroRNA (miRNA) in extracellular vesicles (EVs) has great potential to be a promising marker in liquid biopsy. However, the present EV isolation methods, such as ultracentrifugation, have complicated and long-time operation, which impedes research on EV miRNA. The downstream complex miRNA extraction process will also significantly increase the detection cycle and loss. We first established a simple automated technique to efficiently extract target miRNAs in EVs from plasma based on FeO@TiO beads with high affinity and capture efficiency. We combined a heat-lysis method for quick and simple EV miRNA extraction and detection. The results indicated that our method has more RNA yield than TRIzol or a commercial kit and could complete EV enrichment and miRNA extraction in 30 min. Through the detection of miRNA-21, healthy people and lung cancer patients were distinguished, which verified the possibility of the application in clinical detection. The automated isolation technology for EV miRNA has good repeatability and high throughput, with great application potential in clinical diagnosis.

摘要

细胞外囊泡(EV)中的微小RNA(miRNA)很有潜力成为液体活检中有前景的标志物。然而,目前的EV分离方法,如超速离心,操作复杂且耗时,这阻碍了对EV miRNA的研究。下游复杂的miRNA提取过程也会显著增加检测周期和损失。我们首先基于具有高亲和力和捕获效率的FeO@TiO磁珠建立了一种简单的自动化技术,以有效地从血浆中提取EV中的目标miRNA。我们结合了热裂解方法用于快速简单的EV miRNA提取和检测。结果表明,我们的方法比TRIzol或商业试剂盒具有更高的RNA产量,并且可以在30分钟内完成EV富集和miRNA提取。通过检测miRNA-21,区分了健康人和肺癌患者,这验证了其在临床检测中应用的可能性。EV miRNA的自动化分离技术具有良好的重复性和高通量,在临床诊断中具有巨大的应用潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/537d/9399425/4b61ff768872/fbioe-10-948757-g001.jpg

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