Development of a novel Colorimetric Assay for the rapid diagnosis of Coronavirus disease 2019 from nasopharyngeal samples

Sci Rep. 2024 May 27;14(1):12125. doi: 10.1038/s41598-024-53747-0.

Abstract

Emergence of Coronavirus disease 2019 (COVID-19) pandemic has posed a huge threat to public health. Rapid and reliable test to diagnose infected subjects is crucial for disease spread control. We developed a colorimetric test for COVID-19 detection using a Colorimetric Assay based on thiol-linked RNA modified gold nanoparticles (AuNPs) and oligonucleotide probes. This method was conducted on RNA from 200 pharyngeal swab samples initially tested by Real-Time polymerase chain reaction (RT-PCR) as gold standard. A specific oligonucleotide probe designed based on ORF1ab of COVID-19 was functionalized with AuNPs-probe conjugate. The exposure of AuNP-probe to isolated RNA samples was tested using hybridization. In this comparative study, the colorimetric functionalized AuNPs assay exhibited a detection limit of 25 copies/µL. It was higher in comparison to the RT-PCR method, which could only detect 15 copies/µL. The results demonstrated 100% specificity and 96% sensitivity for the developed method. Herein, we developed an incredibly rapid, simple and cost-effective Colorimetric Assay lasting approximately 30 min which could process considerably higher number of COVID-19 samples compared to the RT-PCR. This AuNP-probe conjugate colorimetric method could be considered the optimum alternatives for conventional diagnostic tools especially in over-populated and/or low-income countries.

Keywords: AuNP-probe conjugate; Coronavirus disease 2019; Diagnosis; Real-Time PCR.

MeSH terms

  • COVID-19 Nucleic Acid Testing / methods
  • COVID-19 Testing / methods
  • COVID-19* / diagnosis
  • COVID-19* / virology
  • Colorimetry* / methods
  • Gold* / chemistry
  • Humans
  • Limit of Detection
  • Metal Nanoparticles* / chemistry
  • Nasopharynx* / virology
  • Oligonucleotide Probes / genetics
  • RNA, Viral* / analysis
  • RNA, Viral* / genetics
  • RNA, Viral* / isolation & purification
  • Real-Time Polymerase Chain Reaction / methods
  • SARS-CoV-2* / genetics
  • SARS-CoV-2* / isolation & purification
  • Sensitivity and Specificity*

Substances

  • Gold
  • RNA, Viral
  • Oligonucleotide Probes