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

Lab Chip. 2023 Oct 24;23(21):4701-4707. doi: 10.1039/d3lc00687e.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Humans
  • Nucleic Acid Amplification Techniques / methods
  • Point-of-Care Systems
  • Point-of-Care Testing*
  • RNA, Viral* / analysis
  • RNA, Viral* / genetics
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcription
  • Sensitivity and Specificity

Substances

  • RNA, Viral