Single-shot multi-channel plasmonic real-time polymerase chain reaction for multi-target point-of-care testing

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Plasmonic nucleic acid amplification tests demand high-throughput and multi-target detection of infectious diseases as well as short turnaround time and small size for point-of-care molecular diagnostics. Here, we report a multi-channel plasmonic real-time reverse-transcription polymerase chain reaction (mpRT-qPCR) assay for ultrafast and on-chip multi-target detection. The mpRT-qPCR system features two pairs of plasmonic thermocyclers for rapid nanostructure-driven amplification and microlens array fluorescence microscopes for in situ multi-color fluorescence quantification. Each channel shows a physical dimension of 32 mm, 75 mm, and 25 mm in width, length, and thickness. The ultrathin microscopes simultaneously capture four different fluorescence images from two PCR chambers of a single cartridge at a single shot exposure per PCR cycle of four different excitation light sources. The experimental results demonstrate a single assay result of high-throughput amplification and multi-target quantification for RNA-dependent RNA polymerase, nucleocapsid, and human ribonuclease P genes in SARS-CoV-2 RNA detection. The mpRT-PCR increases the number of tests four times over the single RT-PCR and exhibits a short detection time of 15 min for the four RT-PCR reactions. This point-of-care molecular diagnostic platform can reduce false negative results in clinical applications of virus detection and decentralize healthcare facilities with limited infrastructure. A multi-channel plasmonic real-time reverse-transcription polymerase chain reaction system is used to demonstrate high-throughput and multi-target detection of the SARS-CoV-2 virus with four simultaneous reactions in a single assay within 15 min.
Publisher
ROYAL SOC CHEMISTRY
Issue Date
2023-10
Language
English
Article Type
Article
Citation

LAB ON A CHIP, v.23, no.21, pp.4701 - 4707

ISSN
1473-0197
DOI
10.1039/d3lc00687e
URI
http://hdl.handle.net/10203/314296
Appears in Collection
BiS-Journal Papers(저널논문)
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