Specific Detection of Influenza A and B Viruses by CRISPR-Cas12a-Based Assay

Bum Ju Park, Man Seong Park, Jae Myun Lee, Yoon Jae Song

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

A rapid and accurate on-site diagnostic test for pathogens including influenza viruses is critical for preventing the spread of infectious diseases. Two types of influenza virus, A and B cause seasonal flu epidemics, whereas type A can cause influenza pandemics. To specifically detect influenza A (IAV) and B (IBV) viruses, we developed a clustered, regularly interspaced, short palindromic repeats (CRISPR) and CRISPR-associated (Cas) system-based assay. By coupling reverse transcription recombinase polymerase amplification (RT-RPA) and reverse transcription loop-mediated isothermal amplification (RT-LAMP), a CRISPR-Cas12a DNA endonuclease-targeted CRISPR trans-reporter (DETECTR) detected IAV and IBV titers as low as 1 × 100 plaque forming units (PFUs) per reaction without exhibiting cross-reactivity. Only 75 to 85 min were required to detect IAV and IBV, depending on isothermal nucleic acid amplification methods, and results were verified using a lateral flow strip assay that does not require additional analytic equipment. Taken together, our findings establish RT-RPA and RT-LAMP-coupled DETECTR-based diagnostic tests for rapid, specific and high-sensitivity detection of IAV and IBV using fluorescence and lateral flow assays. The diagnostic test developed in this study can be used to distinguish IAV and IBV infections, a capability that is necessary for monitoring and preventing the spread of influenza epidemics and pandemics.

Original languageEnglish
JournalBiosensors
Volume11
Issue number3
DOIs
Publication statusPublished - 2021 Mar 19

Keywords

  • CRISPR-Cas12a
  • DETECTR
  • diagnosis
  • influenza virus

ASJC Scopus subject areas

  • Clinical Biochemistry

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