LAMP-Coupled CRISPR–Cas12a Module for Rapid and Sensitive Detection of Plant DNA Viruses

Ahmed Mahas, Norhan Hassan, Rashid Aman, Tin Maršić, Qiaochu Wang, Zahir Ali, Magdy M. Mahfouz

Research output: Contribution to journalArticlepeer-review

54 Scopus citations

Abstract

One important factor for successful disease management is the ability to rapidly and accurately identify the causal agent. Plant viruses cause severe economic losses and pose a serious threat to sustainable agriculture. Therefore, optimization of the speed, sensitivity, feasibility, portability, and accuracy of virus detection is urgently needed. Here, we developed a clustered regularly interspaced short palindromic repeats (CRISPR)-based nucleic acid diagnostic method utilizing the CRISPR–Cas12a system for detecting two geminiviruses, tomato yellow leaf curl virus (TYLCV) and tomato leaf curl New Delhi virus (ToLCNDV), which have single-stranded DNA genomes. Our assay detected TYLCV and ToLCNDV in infected plants with high sensitivity and specificity. Our newly developed assay can be performed in ~1 h and provides easy-to-interpret visual readouts using a simple, low-cost fluorescence visualizer, making it suitable for point-of-use applications.
Original languageEnglish (US)
Pages (from-to)466
JournalViruses
Volume13
Issue number3
DOIs
StatePublished - Mar 12 2021

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

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