Choi, Woongsun Yeom, Sang Yun Kim, Junsun Jung, Seungwon Jung, Seungho Shim, Tae Soup Kim, Sang Kyung Kang, Ji Yoon Lee, Soo Hyun Cho, Il-Joo Choi, Jungkyu Choi, Nakwon 2024-01-19T23:03:59Z 2024-01-19T23:03:59Z 2021-09-03 2018-03-15 0956-5663 https://pubs.kist.re.kr/handle/201004/121597 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. English ELSEVIER ADVANCED TECHNOLOGY REAL-TIME PCR QRT-PCR WEST NILE MICRORNA EXPRESSION ENCEPHALITIS-VIRUS MICROARRAY BIOMARKERS RNA IDENTIFICATION ASSAY Hydrogel micropost-based qPCR for multiplex detection of miRNAs associated with Alzheimer's disease Article 10.1016/j.bios.2017.10.039 1 BIOSENSORS & BIOELECTRONICS, v.101, pp.235 - 244 BIOSENSORS & BIOELECTRONICS 101 235 244 scie scopus 000418982600032 2-s2.0-85032296060 Biophysics Biotechnology & Applied Microbiology Chemistry, Analytical Electrochemistry Nanoscience & Nanotechnology Biophysics Biotechnology & Applied Microbiology Chemistry Electrochemistry Science & Technology - Other Topics Article REAL-TIME PCR QRT-PCR WEST NILE MICRORNA EXPRESSION ENCEPHALITIS-VIRUS MICROARRAY BIOMARKERS RNA IDENTIFICATION ASSAY Polyethylene glycol diacrylate (PEGDA) Hydrogel micropost Multiplex qPCR miRNA Alzheimer&apos s disease Plastic substrate