Simultaneous detection of multiple exosomal microRNAs for exosome screening based on rolling circle amplification
Simultaneous detection of multiple exosomal microRNAs for exosome screening based on rolling circle amplification
复制标题
基于滚环扩增同时检测多种外泌体microRNA用于外泌体筛选
DOI：
10.1088/1361-6528/abc7d4
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发表时间：
2020-11
期刊：
影响因子：
3.5
通讯作者：
Yiping Cui
中科院分区：
文献类型：
--
作者：
Zhile Wang;Shenfei Zong;Yun Liu;Ziting Qian;Kai Zhu;Zhaoyan Yang;Zhuyuan Wang;Yiping Cui
外泌体microRNAs （miRNAs）作为癌症预测和预后的生物标志物受到了广泛关注。mirna的分析在癌症的有效诊断中起着关键作用。然而，由于它们的同源性和低丰度，特别是在外泌体中，同时定量多个mirna是具有挑战性的。在这里，我们开发了一种利用滚动环扩增（RCA）对多个外泌体mirna进行灵敏检测的方法。与传统方法相比，该方法既具有多路感知能力，又具有RCA算法简单的优点。具体来说，来自不同细胞系的多个外泌体mirna通过RCA同时复制，并使用设计的分子信标（mb）进行检测。我们选择miRNA-21、miRNA-122和miRNA-155作为靶点，它们在癌症中过表达。MB的归一化荧光强度表示这些mirna的相对浓度。获得的相对mirna表达水平可用于区分乳腺癌外泌体和正常外泌体。如果检测到的外泌体mirna的种类足够丰富，则mirna的浓度比基本可以表明相应的外泌体，可以实现外泌体的筛选。这种外泌体miRNA谱分析和外泌体筛选可以辅助癌症诊断，具有很好的临床应用前景。
Exosomal microRNAs (miRNAs) have attracted great attention as predictive and prognostic biomarkers of cancer. Profiling of miRNAs plays a key role in the effective diagnosis of cancers. However, simultaneous quantification of multiple miRNAs is challenging due to their homology and low abundance especially in exosomes. Here, we developed a sensitive detection method for multiple exosomal miRNAs with the help of rolling circle amplification (RCA). In contrast of the traditional ways, this method takes the advantages of both the multiplex sensing ability and the simplicity of RCA. Specifically, multiple exosomal miRNAs from different cell lines were replicated simultaneously through RCA and detected using designed molecular beacons (MBs). miRNA-21, miRNA-122 and miRNA-155 were chosen as the targets, which are overexpressed in cancers. Normalized fluorescence intensities of MB were used to imply the relative concentrations of these miRNAs. The obtained relative miRNAs expression levels could be used to distinguish the breast cancer exosome from normal one. If the varieties of the detected exosomal miRNAs are abundant enough, the concentration ratios of miRNAs could basically indicate the corresponding exosome and exosome screening could be realized. Such exosomal miRNA profiling and exosome screening can assist cancer diagnosis, which is promising in clinical application.
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影响因子：
7.4
作者：
Yaokun Xia;Liangliang Wang;Juan Li;Xiangqi Chen;Jianming Lan;An Yan;Yun Lei;Sheng Yang;
通讯作者：
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影响因子：
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作者：
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通讯作者：
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DOI：
10.1007/978-1-61779-163-5
发表时间：
2011
期刊：
--
影响因子：
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作者：
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通讯作者：
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影响因子：
16.6
作者：
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通讯作者：
Yan, Hao
影响因子：
8.9
作者：
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通讯作者：
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