Label-free Colorimetric Detection of Viral RNA Based on Clustered Regularly Interspaced Short Palindromic Repeats and Gold Nanoparticles with a Portable Device

Langmuir. 2024 Jun 4;40(22):11534-11540. doi: 10.1021/acs.langmuir.4c00657. Epub 2024 May 17.

Abstract

Viral infections, such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), are some of the most dangerous threats to humans. SARS-CoV-2 has caused a global pandemic, highlighting the unprecedented demand for rapid and portable diagnostic methods. To meet these requirements, we designed a label-free colorimetric platform that combines the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated proteins (Cas) 12a system for naked-eye detection (named LFP). This method utilizes reverse transcription loop-mediated isothermal amplification (RT-LAMP) and the trans-cleavage activity of the CRISPR/Cas12a system to increase the sensitivity and specificity of the reaction. This platform can detect as few as 4 copies/μL of RNA and produces no false positive results when tested against the influenza virus. To better meet the requirements of point-of-care (POC) detection, we developed a portable device that can be applied in resource-poor and densely populated regions. The LFP assay holds great potential for application in resource-limited settings, and the label-free gold nanoparticle (AuNPs) probe can reduce costs, making it suitable for large-scale screening. We expect that the LFP assay will be promising for the POC screening of COVID-19.

MeSH terms

  • COVID-19 / diagnosis
  • COVID-19 / virology
  • CRISPR-Cas Systems
  • Clustered Regularly Interspaced Short Palindromic Repeats
  • Colorimetry* / instrumentation
  • Colorimetry* / methods
  • Gold* / chemistry
  • Humans
  • Metal Nanoparticles* / chemistry
  • Molecular Diagnostic Techniques
  • Nucleic Acid Amplification Techniques* / instrumentation
  • Nucleic Acid Amplification Techniques* / methods
  • RNA, Viral* / analysis
  • RNA, Viral* / genetics
  • SARS-CoV-2* / genetics
  • SARS-CoV-2* / isolation & purification

Substances

  • Gold
  • RNA, Viral

Supplementary concepts

  • LAMP assay