Crafting a streamlined isothermal amplification assay for sensitive and rapid nucleic acid detection of glioblastoma

Yu He Liu, Yi Chien Chen, Li Hsien Ho, Zhi Yang Wei, Chia Hung Chen, Yun Hsien Chung, I. Hsuan Chou, Tao Yun Yen, Wei Chun Lan, Hsien Wen Yao, Peng Wei Hsu, Chen Han Huang, Hsing Ying Lin

研究成果: 書貢獻/報告類型會議論文篇章同行評審


Development of liquid biopsies for non-invasive tumor characterization techniques shows a great promise for real-time monitoring of tumor recurrence, progression, and treatment response. To better assess the various tumor evolution, cellular heterogeneity, consequent drug-resistance mechanisms, it is critical to screen specific proteins and nucleic acids from biofluid-derived extracellular vesicles (EVs). We are developing a rapid integrated ultra-sensitive digital bead-based sensor enabling evaluation of molecular profiling of EVs in glioblastoma (GBM). The tumor relevant nucleic acid information in EVs is enriched by antibody-specific magnetic beads and amplified by reverse transcription recombinase polymerase amplification using the CRISPR/Cas13a system with around twenty-five thousand droplets. Detection results are analyzed by a miniaturized portable imaging device. The niches of our developing technique include: (a) the streamlined sample preparation process and miniaturized detection system enable the sensitive detection of important mRNAs, EGFR, EGFRvIII, IDH1wt, IDH1 R132H and GAPDH, in EVs within 1.5 hours; (b) the fluorescent signal-to-noise ratio (SNR) in each micrometer sized droplet is significantly enhanced via using a simple Cas13a assay and triggering greater than 104 turnovers of fluorescent reporters on a high-throughput microfluidic chip; (c) real-time image data are transferred to a cloud-based server and classified using several trained YOLOv5 models; (d) the RNAomics results are displayed on the Raspberry Pi touchscreen. This approach improves the speed of GBM diagnosis and treatment decision-making in using liquid biopsy.

主出版物標題Biomedical Imaging and Sensing Conference
編輯Toyohiko Yatagai
出版狀態已出版 - 2023
事件Biomedical Imaging and Sensing Conference 2023 - Yokohama, Japan
持續時間: 17 4月 202321 4月 2023


名字Proceedings of SPIE - The International Society for Optical Engineering


???event.eventtypes.event.conference???Biomedical Imaging and Sensing Conference 2023


深入研究「Crafting a streamlined isothermal amplification assay for sensitive and rapid nucleic acid detection of glioblastoma」主題。共同形成了獨特的指紋。