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Hydrogel micropost-based qPCR for multiplex detection of miRNAs associated with Alzheimer's disease

Authors
Choi, WoongsunYeom, Sang YunKim, JunsunJung, SeungwonJung, SeunghoShim, Tae SoupKim, Sang KyungKang, Ji YoonLee, Soo HyunCho, Il-JooChoi, JungkyuChoi, Nakwon
Issue Date
15-3월-2018
Publisher
ELSEVIER ADVANCED TECHNOLOGY
Keywords
Polyethylene glycol diacrylate (PEGDA); Hydrogel micropost; Multiplex qPCR; miRNA; Alzheimer' s disease; Plastic substrate
Citation
BIOSENSORS & BIOELECTRONICS, v.101, pp.235 - 244
Indexed
SCIE
SCOPUS
Journal Title
BIOSENSORS & BIOELECTRONICS
Volume
101
Start Page
235
End Page
244
URI
https://scholar.korea.ac.kr/handle/2021.sw.korea/76726
DOI
10.1016/j.bios.2017.10.039
ISSN
0956-5663
Abstract
Quantitative polymerase chain reaction (qPCR) renders profiling of genes of interest less time-consuming and cost-effective. Recently, multiplex profiling of miRNAs has enabled identifying or investigating predominant miRNAs for various diseases such as cancers and neurodegenerative diseases. Conventional multiplex qPCR technologies mostly use colorimetric measurements in solution phase, yet not only suffer from limited multiplexing capacity but also require target-screening processes due to non-specific binding between targets and primers. Here, we present hydrogel micropost-based qPCR for multiplex detection of miRNAs associated with Alzheimer's disease (AD). Our methodology promises two key advantages compared with the conventional solution-based PCR: 1) nearly no non-specific crosstalks between targets and primers, and 2) practically valuable multiplexing by spatial encoding within a single microchamber. Specifically, we immobilized hydrogel micro posts (similar to 400 mu m in diameter) within commercially available polycarbonate PCR chips by multi-step ultraviolet (UV, 365 nm) exposure. We optimized this photoimmobilization for thermal cycles of PCR as well. Acrylated forward primers incorporated in polyethylene glycol diacrylate (PEGDA) posts played a crucial role to confine fluorescent signal of cDNA amplification within the PEGDA hydrogel. To demonstrate the potential of our platform, we successfully verified multiplex detection of five miRNAs, which were reported to be highly correlated with AD, from a complex buffer of human plasma.
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공과대학 (화공생명공학과)
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