한빛사
>
한빛사논문
한빛사논문
- 조회185
Biosens. Bioelectron., Available online 17 February 2025, 117277 | https://doi.org/10.1016/j.bios.2025.117277
FlexPCR: A Streamlined Multiplexed Digital mRNA Quantification Platform with Universal Primers and Limited Fluorescence Channels
Joon Soo Park 1, Jiumei Hu 2, Liben Chen 2, Tza-Huei Wang 1,2,3
1Department of Biomedical Engineering, Johns Hopkins University, Baltimore, MD 21218, USA
2Department of Mechanical Engineering, Johns Hopkins University, Baltimore, MD 21218, USA
3Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, 21218, USA
Corresponding author : Tza-Huei Wang
1Department of Biomedical Engineering, Johns Hopkins University, Baltimore, MD 21218, USA
2Department of Mechanical Engineering, Johns Hopkins University, Baltimore, MD 21218, USA
3Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, 21218, USA
Corresponding author : Tza-Huei Wang
Abstract
Accurate quantification of multiple messenger RNA (mRNA) targets is essential for biomedical research and disease diagnosis. Current PCR-based methods for mRNA analysis are limited by the number of fluorescent labels and the complexities associated with multiple target-specific primers, leading to amplification bias and limited multiplexing capability. Here, we introduce Fluorescence-coding extension PCR (FlexPCR), a novel digital PCR-based assay that overcomes these limitations by employing a universal primer and probe strategy in conjugation with oligo extension. This method generates unique fluorescence-coded PCR templates for each mRNA target, enabling multiplexed detection using minimal fluorescence channels. FlexPCR simplifies assay design, reduces non-specific amplification, and enhances quantification accuracy. We demonstrate the efficacy by quantifying seven immune response mRNAs using only two fluorescence colors in various human total RNA samples. The results correlate strongly with gold-standard single-plex RT-qPCR, validating the accuracy of our method. FlexPCR offers a streamlined and scalable approach for multiplexed mRNA quantification with broad applications in gene expression analysis and molecular diagnostics.
논문정보
- Research article
- 2025년 02월 (BRIC 등록일 2025-02-18)
- 국외 연구진
- 바이오·의료융합 > 바이오센싱 및 나노바이오물질
댓글 작성 로그인
팝업 닫기관련 링크
연구자 키워드
관련분야 연구자보기
소속기관 논문보기
관련분야 논문보기
해당논문 저자보기
