Application of the amplification-free SERS-based CRISPR/Cas12a platform in the identification of SARS-CoV-2 from clinical samples

J Nanobiotechnology. 2021 Sep 8;19(1):273. doi: 10.1186/s12951-021-01021-0.

Abstract

The control of contagious or refractory diseases requires early, rapid diagnostic assays that are simple, fast, and easy-to-use. Here, easy-to-implement CRISPR/Cas12a-based diagnostic platform through Raman transducer generated by Raman enhancement effect, term as SERS-CRISPR (S-CRISPR), are described. The S-CRISPR uses high-activity noble metallic nanoscopic materials to increase the sensitivity in the detection of nucleic acids, without amplification. This amplification-free platform, which can be performed within 30-40 min of incubation time, is then used for detection of SARS-CoV-2 derived nucleic acids in RNA extracts obtained from nasopharyngeal swab specimens (n = 112). Compared with the quantitative reverse transcription polymerase chain reaction (RT-qPCR), the sensitivity and specificity of S-CRISPR reaches 87.50% and 100%, respectively. In general, the S-CRISPR can rapidly identify the RNA of SARS-CoV-2 RNA without amplification and is a potential strategy for nucleic acid point of care test (POCT).

Keywords: Biosensors; COVID-19; CRISPR; Cas12a; SARS-CoV-2; Sliver nanoparticle; Surface-enhanced Raman scattering.

MeSH terms

  • COVID-19 / diagnosis
  • COVID-19 / virology
  • CRISPR-Cas Systems / genetics*
  • Gene Expression Regulation, Fungal
  • Genes, Viral
  • Humans
  • Nucleic Acid Amplification Techniques*
  • RNA, Viral / analysis
  • SARS-CoV-2 / genetics*
  • SARS-CoV-2 / isolation & purification*
  • Sensitivity and Specificity
  • Spectrum Analysis, Raman*

Substances

  • RNA, Viral