Vigilant: An Engineered VirD2-Cas9 Complex for Lateral Flow Assay-Based Detection of SARS-CoV2

Tin Marsic, Zahir Ali, Muhammad Tehseen, Ahmed Mahas, Samir Hamdan, Magdy M. Mahfouz

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

Rapid, sensitive, and specific point-of-care testing for pathogens is crucial for disease control. Lateral flow assays (LFAs) have been employed for nucleic acid detection, but they have limited sensitivity and specificity. Here, we used a fusion of catalytically inactive SpCas9 endonuclease and VirD2 relaxase for sensitive, specific nucleic acid detection by LFA. In this assay, the target nucleic acid is amplified with biotinylated oligos. VirD2-dCas9 specifically binds the target sequence via dCas9 and covalently binds to a FAM-tagged oligonucleotide via VirD2. The biotin label and FAM tag are detected by a commercially available LFA. We coupled this system, named Vigilant (VirD2-dCas9 guided and LFA-coupled nucleic acid test), to reverse transcription-recombinase polymerase amplification to detect SARS-CoV2 in clinical samples. Vigilant exhibited a limit of detection of 2.5 copies/μL, comparable to CRISPR-based systems, and showed no cross-reactivity with SARS-CoV1 or MERS. Vigilant offers an easy-to-use, rapid, cost-effective, and robust detection platform for SARS-CoV2.
Original languageEnglish (US)
JournalNano Letters
DOIs
StatePublished - Apr 12 2021

ASJC Scopus subject areas

  • Bioengineering
  • Materials Science(all)
  • Chemistry(all)
  • Mechanical Engineering
  • Condensed Matter Physics

Fingerprint

Dive into the research topics of 'Vigilant: An Engineered VirD2-Cas9 Complex for Lateral Flow Assay-Based Detection of SARS-CoV2'. Together they form a unique fingerprint.

Cite this