Department of Medical Device Management and Research, Samsung Advanced Institute for Health Sciences & Technology, Sungkyunkwan University, 115, Irwon-ro, Gangnam-gu, Seoul, 06355, Republic of Korea; Biomedical Engineering Research Center, Samsung Medical Center, 81, Irwon-ro, Gangnam-gu, Seoul, 06351, Republic of Korea.
Department of Nano-Bio Convergence, Korea Institute of Materials Science (KIMS), Changwon, Gyeongnam, 51508, Republic of Korea.
Biosens Bioelectron. 2021 Jun 15;182:113167. doi: 10.1016/j.bios.2021.113167. Epub 2021 Mar 19.
The current clinically available multiplex molecular diagnostic technologies are difficult to apply to onsite diagnostic platforms due to their large and sophisticated instrumentation, long amplification times and limited number of simultaneous detections. We developed a plasmonic isothermal recombinase polymerase amplification (RPA) array chip for rapid and sensitive multiplex molecular detection. The 3D plasmonic substrate composed of Au nanoparticles (NPs) on dense Au nanopillars (NPOP) showed highly enhanced plasmon-enhanced fluorescence (PEF) of RPA products with long DNA amplicons (~200 bp). The plasmonic 4-plex RPA array chip successfully detected bacterial DNA within 30 min and viral RNA within 40 min; the sensitivity of the plasmonic RPA array chip was comparable to or 10-fold higher than that of the 4-pelx liquid-phase RPA and 4-plex liquid-phase PCR techniques. Additionally, no cross-reactivity was observed. The 4-plex plasmonic RPA array chip was preliminary evaluated using clinical respiratory viral-positive nasopharyngeal swab samples. This approach enables rapid, sensitive and high-multiplex molecular detection and can be used in the realization of a simplified and miniaturized platform for onsite multiplex molecular diagnostics.
目前临床可用的多重分子诊断技术由于其大型复杂的仪器、较长的扩增时间和有限的同时检测数量，难以应用于现场诊断平台。我们开发了一种用于快速灵敏的多重分子检测的等离子体等温重组酶聚合酶扩增（RPA）阵列芯片。由密集 Au 纳米柱（NPOP）上的 Au 纳米颗粒（NPs）组成的 3D 等离子体基底对具有长 DNA 扩增子（~200 bp）的 RPA 产物表现出高度增强的等离子体增强荧光（PEF）。该等离子体 4 重 RPA 阵列芯片成功地在 30 分钟内检测到细菌 DNA，在 40 分钟内检测到病毒 RNA；等离子体 RPA 阵列芯片的灵敏度与 4 重液相 RPA 和 4 重液相 PCR 技术相当，或高 10 倍。此外，没有观察到交叉反应。该 4 重等离子体 RPA 阵列芯片已初步用于临床呼吸道病毒阳性鼻咽拭子样本的检测。该方法能够实现快速、灵敏和高多重分子检测，并可用于简化和小型化现场多重分子诊断平台的实现。
