A rapid and label-free DNA-based interference reduction nucleic acid amplification strategy for viral RNA detection

Feng Chen, Guodong Li, Chun Wu, Wanhe Wang, Dik Lung Ma*, Chung Hang Leung*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)

Abstract

Common reference methods for COVID-19 diagnosis include thermal cycling amplification (e.g. RT-PCR) and isothermal amplification methods (e.g. LAMP and RPA). However, they may not be suitable for direct detection in environmental and biological samples due to background signal interference. Here, we report a rapid and label-free interference reduction nucleic acid amplification strategy (IR-NAAS) that exploits the advantages of luminescent iridium(III) probes, time-resolved emission spectroscopy (TRES) and multi-branch rolling circle amplification (mbRCA). Using IR-NAAS, we established a luminescence approach for diagnosing COVID-19 RNAs sequences RdRp, ORF1ab and N with a linear range of 0.06–6.0 × 105 copies/mL and a detection limit of down to 7.3 × 104 copies/mL. Moreover, the developed method was successfully applied to detect COVID-19 RNA sequences from various environmental and biological samples, such as domestic sewage, and mice urine, blood, feces, lung tissue, throat and nasal secretions. Apart from COVID-19 diagnosis, IR-NAAS was also demonstrated for detecting other RNA viruses, such as H1N1 and CVA10, indicating that this approach has great potential approach for routine preliminary viral detection.

Original languageEnglish
Article number113829
Number of pages9
JournalBiosensors and Bioelectronics
Volume198
DOIs
Publication statusPublished - 15 Feb 2022

Scopus Subject Areas

  • Biotechnology
  • Biophysics
  • Biomedical Engineering
  • Electrochemistry

User-Defined Keywords

  • Coronavirus detection
  • G-quadruplex
  • Iridium(III) complex
  • Rolling circle amplification

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