Sensitive and Easy-Read CRISPR Strip for COVID-19 Rapid Point-of-Care Testing

CRISPR J. 2021 Jun;4(3):392-399. doi: 10.1089/crispr.2020.0138.

Abstract

Rapid and clinically sensitive detection of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) play an important role in the contact tracing and containment of the COVID-19 pandemic. A recently developed field-deployable clustered regularly interspaced short palindromic repeats (CRISPR) detection assay with lateral flow strips shows promise for point-of-care detection of SARS-CoV-2. However, the limit of detection of paper strip-based assays (10-100 copies/μL) is much lower than that of fluorescence-based detection methods. In this study, we developed an easy-readout and sensitive enhanced (ERASE) strip to visualize the results of CRISPR detection and improve the sensitivity to 1 copy/μL with an unambiguous easy-read result. Using 649 clinical samples from blind specimens collected from patients in China, we validated our ERASE assay for SARS-CoV-2 RNA detection with 90.67% positive predictive agreement and 99.21% negative predictive agreement. In conclusion, our study provided a customized CRISPR strip for use in a simple, rapid, ultrasensitive, and highly specific assay for SARS-CoV-2 detection. (Clinical Trial Registration number: 2020-008-01; [2020]IEC(ZD01); PJ-NBEY-2020-009-01; 2020#34).

Publication types

  • Clinical Trial
  • Research Support, Non-U.S. Gov't

MeSH terms

  • COVID-19 / diagnosis*
  • COVID-19 / virology
  • COVID-19 Nucleic Acid Testing / instrumentation*
  • CRISPR-Cas Systems / genetics*
  • Humans
  • Limit of Detection
  • Point-of-Care Testing*
  • Predictive Value of Tests
  • RNA, Viral / genetics
  • RNA, Viral / isolation & purification
  • Reagent Kits, Diagnostic
  • SARS-CoV-2 / genetics
  • SARS-CoV-2 / isolation & purification*

Substances

  • RNA, Viral
  • Reagent Kits, Diagnostic