公开(公告)号：US20240124917A1

公开(公告)日：2024-04-18

申请号：US18546515

申请日：2022-02-15

Applicant: King Abdullah University of Science and Technology

Inventor： Magdy Mahfouz ,  Tin Marsic

IPC: C12Q1/6816 ,  C12N9/22

CPC classification number: C12Q1/6816 ,  C12N9/22 ,  C07K2319/80

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. A fusion of catalytically inactive Cas9 endonuclease and a relaxase for example, VirD2 are used for sensitive, specific nucleic acid detection by LFA. VirD2-dCas9 specifically binds the target nucleic acid sequence via dCas9 and covalently binds to a FAM-tagged oligonucleotide via VirD2. The biotin label and FAM tag are detected using a LFA. This system, termed Vigilant (VirD2-dCas9 guided and LFA-coupled nucleic acid test) is coupled to reverse transcription-recombinase polymerase amplification to detect pathogenic nucleic acid of interest in a sample, it exhibits an impressive limit of detection and shows no cross-reactivity, thus reducing incidents of false positives. Vigilant offers an easy-to-use, rapid, cost-effective, and robust detection platform for SARS-CoV2.