Digital quantification of miRNA directly in plasma using integrated comprehensive droplet digital detection

Kaixiang Zhang, Dong Ku Kang, M. Monsur Ali, Linan Liu, Louai Labanieh, Mengrou Lu, Hamidreza Riazifar, Thi N. Nguyen, Jason A. Zell, Michelle A. Digman, Enrico Gratton, Jinghong Li, Weian Zhao

Research output: Contribution to journalArticle

38 Scopus citations

Abstract

Quantification of miRNAs in blood can be potentially used for early disease detection, surveillance monitoring and drug response evaluation. However, quantitative and robust measurement of miRNAs in blood is still a major challenge in large part due to their low concentration and complicated sample preparation processes typically required in conventional assays. Here, we present the 'Integrated Comprehensive Droplet Digital Detection' (IC 3D) system where the plasma sample containing target miRNAs is encapsulated into microdroplets, enzymatically amplified and digitally counted using a novel, high-throughput 3D particle counter. Using Let-7a as a target, we demonstrate that IC 3D can specifically quantify target miRNA directly from blood plasma at extremely low concentrations ranging from 10s to 10000 copies per mL in ≤3 hours without the need for sample processing such as RNA extraction. Using this new tool, we demonstrate that target miRNA content in colon cancer patient blood is significantly higher than that in healthy donor samples. Our IC 3D system has the potential to introduce a new paradigm for rapid, sensitive and specific detection of low-abundance biomarkers in biological samples with minimal sample processing.

Original languageEnglish (US)
Pages (from-to)4217-4226
Number of pages10
JournalLab on a Chip
Volume15
Issue number21
DOIs
StatePublished - 2015

All Science Journal Classification (ASJC) codes

  • Bioengineering
  • Biochemistry
  • Chemistry(all)
  • Biomedical Engineering

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  • Cite this

    Zhang, K., Kang, D. K., Ali, M. M., Liu, L., Labanieh, L., Lu, M., Riazifar, H., Nguyen, T. N., Zell, J. A., Digman, M. A., Gratton, E., Li, J., & Zhao, W. (2015). Digital quantification of miRNA directly in plasma using integrated comprehensive droplet digital detection. Lab on a Chip, 15(21), 4217-4226. https://doi.org/10.1039/c5lc00650c