Ultra-sensitive and high-throughput CRISPR-p owered COVID-19 diagnosis

Zhen Huang, Di Tian, Yang Liu, Zhen Lin, Christopher J. Lyon, Weihua Lai, Dahlene Fusco, Arnaud Drouin, Xiaoming Yin, Tony Hu, Bo Ning

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

Recent research suggests that SARS-CoV-2-infected individuals can be highly infectious while asymptomatic or pre-symptomatic, and that an infected person may infect 5.6 other individuals on average. This situation highlights the need for rapid, sensitive SARS-CoV-2 diagnostic assays capable of high-throughput operation that can preferably utilize existing equipment to facilitate broad, large-scale screening efforts. We have developed a CRISPR-based assay that can meet all these criteria. This assay utilizes a custom CRISPR Cas12a/gRNA complex and a fluorescent probe to detect target amplicons produced by standard RT-PCR or isothermal recombinase polymerase amplification (RPA), to allow sensitive detection at sites not equipped with real-time PCR systems required for qPCR diagnostics. We found this approach allowed sensitive and robust detection of SARS-CoV-2 positive samples, with a sample-to-answer time of ~50 min, and a limit of detection of 2 copies per sample. CRISPR assay diagnostic results obtained nasal swab samples of individuals with suspected COVID-19 cases were comparable to paired results from a CDC-approved quantitative RT-PCR (RT-qPCR) assay performed in a state testing lab, and superior to those produced by same assay in a clinical lab, where the RT-qPCR assay exhibited multiple invalid or inconclusive results. Our assay also demonstrated greater analytical sensitivity and more robust diagnostic performance than other recently reported CRISPR-based assays. Based on these findings, we believe that a CRISPR-based fluorescent application has potential to improve current COVID-19 screening efforts.

Original languageEnglish (US)
Article number112316
JournalBiosensors and Bioelectronics
Volume164
DOIs
StatePublished - Sep 15 2020

Keywords

  • COVID-19
  • CRISPR
  • Fluorescent detection
  • Highly sensitive
  • Molecular diagnosis
  • SARS-CoV-2

ASJC Scopus subject areas

  • Biotechnology
  • Biophysics
  • Biomedical Engineering
  • Electrochemistry

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  • Cite this

    Huang, Z., Tian, D., Liu, Y., Lin, Z., Lyon, C. J., Lai, W., Fusco, D., Drouin, A., Yin, X., Hu, T., & Ning, B. (2020). Ultra-sensitive and high-throughput CRISPR-p owered COVID-19 diagnosis. Biosensors and Bioelectronics, 164, [112316]. https://doi.org/10.1016/j.bios.2020.112316