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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 
 BIOSENSORS, Vol.11(3) : 88, 2021-03 
Journal Title
Issue Date
CRISPR-Cas Systems* ; Clustered Regularly Interspaced Short Palindromic Repeats ; Herpesvirus 1, Cercopithecine ; Humans ; Influenza A virus / isolation & purification* ; Influenza B virus / isolation & purification* ; Influenza, Human / diagnosis ; Molecular Diagnostic Techniques ; Nucleic Acid Amplification Techniques ; Pandemics ; Reverse Transcription ; Sensitivity and Specificity
CRISPR-Cas12a ; DETECTR ; diagnosis ; influenza virus
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.
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1. College of Medicine (의과대학) > Dept. of Microbiology (미생물학교실) > 1. Journal Papers
Yonsei Authors
Lee, Jae Myun(이재면) ORCID logo https://orcid.org/0000-0002-5273-3113
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