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

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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