Rapid and Sensitive MicroRNA Detection with Laminar Flow-Assisted Dendritic Amplification on Power-Free Microfluidic Chip

Hideyuki Arata,Kazuo Hosokawa,Hiroshi Komatsu,Mizuo Maeda,Wei-Chun Chin

doi:10.1371/journal.pone.0048329

Abstract

Detection of microRNAs, small noncoding single-stranded RNAs, is one of the key topics in the new generation of cancer research because cancer in the human body can be detected or even classified by microRNA detection. This report shows rapid and sensitive microRNA detection using a power-free microfluidic device, which is driven by degassed poly(dimethylsiloxane), thus eliminating the need for an external power supply. MicroRNA is detected by sandwich hybridization, and the signal is amplified by laminar flow-assisted dendritic amplification. This method allows us to detect microRNA of specific sequences at a limit of detection of 0.5 pM from a 0.5 µL sample solution with a detection time of 20 min. Together with the advantages of self-reliance of this device, this method might contribute substantially to future point-of-care early-stage cancer diagnosis.

Highlights

MicroRNAs are short, highly conserved non-proteincoding single-stranded RNAs of typically 18 to 24 bases
The target miRNA was efficiently detected by sandwich hybridization followed by laminar flow-assisted dendritic amplification (LFDA)
The limit of detection (LOD) was 0.5 pM from a 0.5 mL sample solution, which corresponds to 0.25 amol

Summary

Introduction

MicroRNAs (miRNAs) are short, highly conserved non-proteincoding single-stranded RNAs of typically 18 to 24 bases. Highly sensitive miRNA detection will open a new field of early-stage cancer diagnosis [2,3,4,5,6,7,8,9] Various techniques such as quantitative polymerase chain reaction (qPCR), deep sequencing, and oligo microarrays are the major technical methods for miRNA profiling with their own strengths and drawbacks [10]. Considering point-of-care (POC) diagnosis, there are several additional requirements besides high sensitivity, such as a short detection time and small sample volume These detection processes should be completed on a portable and compact device, which does not require huge equipment or trained personal for operation. Due to these specific requirements, none of the current techniques meets all the requirements for POC diagnosis

Methods

Results

Conclusion