Absolute quantification of lung cancer related microRNA by droplet digital PCR

Ping Wang; Fengxiang Jing; Gang Li; Zhenhua Wu; Zule Cheng; Jishen Zhang; Honglian Zhang; Chunping Jia; Qinghui Jin; Hongju Mao; Jianlong Zhao

doi:10.1016/j.bios.2015.07.048

Absolute quantification of lung cancer related microRNA by droplet digital PCR

Biosens Bioelectron. 2015 Dec 15:74:836-42. doi: 10.1016/j.bios.2015.07.048. Epub 2015 Jul 22.

Authors

Affiliations

¹ State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Science, Shanghai 200050, China; University of Chinese Academy of Sciences, Beijing 100039, China.
² State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Science, Shanghai 200050, China.
³ State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Science, Shanghai 200050, China; School of Optoelectronic Engineering, Chongqing University, Chongqing 400044, China.
⁴ State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Science, Shanghai 200050, China. Electronic address: hjmao@mail.sim.ac.cn.
⁵ State Key Laboratory of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Science, Shanghai 200050, China. Electronic address: jlzhao@mail.sim.ac.cn.

PMID: 26232679
DOI: 10.1016/j.bios.2015.07.048

Abstract

Digital polymerase chain reaction (digital PCR) enables the absolute quantification of nucleic acids through the counting of single molecules, thus eliminating the need for standard curves or endogenous controls. In this study, we developed a droplet digital PCR (ddPCR) system based on an oil saturated PDMS (OSP) microfluidic chip platform for quantification of lung cancer related microRNA (miRNA). The OSP chip was made with PDMS and was oil saturated to constrain oil swallow and maintain the stability of droplets. Two inlets were designed for oil and sample injection with a syringe pump at the outlet. Highly uniform monodisperse water-in-oil emulsion droplets to be used for subsequent detection and analysis were generated at the cross section of the channel. We compared miRNA quantification by the ddPCR system and quantitative real-time PCR (qPCR) to demonstrate that the ddPCR system was superior to qPCR both in its detection limit and smaller fold changes measurement. This droplet PCR system provides new possibilities for highly sensitive and efficient detection of cancer-related genes.

Keywords: Digital PCR; Lung cancer; Microfluidic chip; miRNA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Biosensing Techniques*
Humans
Limit of Detection
Lung Neoplasms / genetics*
Lung Neoplasms / pathology
MicroRNAs / genetics
MicroRNAs / isolation & purification*
Microfluidics
Neoplasms / diagnosis*
Polymerase Chain Reaction

Substances

MicroRNAs