4.7 Article

Microfluidic exponential rolling circle amplification for sensitive microRNA detection directly from biological samples

期刊

SENSORS AND ACTUATORS B-CHEMICAL
卷 279, 期 -, 页码 447-457

出版社

ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2018.09.121

关键词

Microfluidic exponential rolling circle; amplification (MERCA); microRNA detection; Raw cell lysate; Cell-derived exosomes; Complex biological samples

资金

  1. NIH [1R21EB024101, 1R21CA186846, 1R33CA214333, 1R21CA207816, P20GM103638]
  2. Kansas IDeA Network of Biomedical Research Excellence (K-INBRE) from NIH/NIGMS [P20GM103418]

向作者/读者索取更多资源

There is an urgent need of sensitive bioanalytical platforms for sensitive and precise quantification of low-abundance microRNA targets in complex biological samples, including liquid biopsies of tumors. Many of current miRNA biosensing methods require laborious sample pretreatment procedures, including extraction of total RNA, which largely limits their biomedical and clinical applications. Herein we developed an integrated Microfluidic Exponential Rolling Circle Amplification (MERCA) platform for sensitive and specific detection of microRNAs directly in minimally processed samples. The MERCA system integrates and streamlines solid-phase miRNA isolation, miRNA-adapter ligation, and a dual-phase exponential rolling circle amplification (eRCA) assay in one analytical workflow. By marrying the advantages of microfluidics in leveraging bioassay performance with the high sensitivity of eRCA, our method affords a remarkably low limit of detection at < 10 zeptomole levels, with the ability to discriminate single-nucleotide difference. Using the MERCA chip, we demonstrated quantitative detection of miRNAs in total RNA, raw cell lysate, and cell-derived exosomes. Comparing with the parallel TaqMan RT-qPCR measurements verified the adaptability of the MERCA system for detection of miRNA biomarkers in complex biological materials. In particular, high sensitivity of our method enables direct detection of low-level exosomal miRNAs in as few as 2 x 10(6) exosomes. Such analytical capability immediately addresses the unmet challenge in sample consumption, a key setback in clinical development of exosome-based liquid biopsies. Therefore, the MERCA would provide a useful platform to facilitate miRNA analysis in broad biological and clinical applications.

作者

我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。

评论

主要评分

4.7
评分不足

次要评分

新颖性
-
重要性
-
科学严谨性
-
评价这篇论文

推荐

暂无数据
暂无数据