One-Step RT-PCR for Detection of Micrornas in Exosomes Using Droplet Microfluidics

Binbin Cui, Chunchen Liu, S. Yao
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引用次数: 4

Abstract

The dysregulation of the microRNAs, small noncoding RNA around 22 bases, accounts for a wide range of diseases including cancer. However, current methods technologies for of the microRNA the detection and analysis of microRNA are limited to the analysis of bulk exosomes, which fails to depict the heterogeneity of the exosome population as individual vesicles. In this work, we developed a reverse transcription polymerase chain reaction (RT-PCR) protocol on a droplet microfluidic platform. We employed the synthetic hsa-miR-21-5p and exosomes extracted from lung cancer patients in the proof-of-concept experiments. The results indicated that our method can detect as low as a single molecule of microRNA per droplet, which is promising for single exosome microRNA assays.
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利用微滴微流体一步法RT-PCR检测外泌体中的微rna
microrna是一种约22个碱基的小非编码RNA,其失调是包括癌症在内的多种疾病的原因。然而,目前用于检测和分析microRNA的方法技术仅限于对散装外泌体的分析,无法描述外泌体群体作为单个囊泡的异质性。在这项工作中,我们在液滴微流控平台上开发了一种逆转录聚合酶链反应(RT-PCR)方案。我们将合成的hsa-miR-21-5p和从肺癌患者中提取的外泌体用于概念验证实验。结果表明,我们的方法可以检测到每滴低至单个分子的microRNA,这对于单个外泌体microRNA的检测是有希望的。
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