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

• DOI: 10.1039/c5lc00650c
• Publication Date: 2015-09-14T08:01:26Z
• Authors: 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
• Keywords: 0301 basic medicine; 610; 600; Analytical Chemistry; 4.1 Discovery and preclinical testing of markers and technologies; 3. Good health; Systems Integration; MicroRNAs; 03 medical and health sciences; Engineering; Chemical sciences; Limit of Detection; Lab-On-A-Chip Devices; Chemical Sciences; Colonic Neoplasms; Genetics; Humans; Generic health relevance; Blood Chemical Analysis; Biotechnology; Cancer

Quantification of miRNAs in blood can be potentially used for early disease detection, surveillance monitoring and drug response evaluation. However, quantitative robust measurement is still a major challenge large part due to their low concentration complicated sample preparation processes typically required conventional assays. Here, we present the 'Integrated Comprehensive Droplet Digital Detection' (IC 3D) system where plasma containing target encapsulated into microdroplets, enzymatically amplified digitally counted using novel, high-throughput 3D particle counter. Using Let-7a as target, demonstrate that IC specifically quantify miRNA directly from at extremely concentrations ranging 10s 10 000 copies per mL ≤3 hours without need processing such RNA extraction. this new tool, content colon cancer patient significantly higher than healthy donor samples. Our has potential introduce paradigm rapid, sensitive specific detection low-abundance biomarkers biological samples with minimal processing.