CRISPR‐Cas12a assisted specific detection of mpox virus

Author:

Singh Mandeep1,Misra Chitra Seetharam1,Bindal Gargi12,Rangu Shyam Sunder1,Rath Devashish12ORCID

Affiliation:

1. Applied Genomics Section Bhabha Atomic Research Centre Mumbai India

2. Homi Bhabha National Institute Mumbai India

Abstract

AbstractMpox virus, a member of genus Orthopoxvirus, causes rash and flu‐like symptoms in humans. In the recent global outbreak, it was reported from several geographical areas that have not historically reported mpox. Point of care, sensitive and specific mpox diagnostic assays are critical in checking the spread of the disease. We have developed a clustered regularly interspaced short palindromic repeats associated Cas12a nuclease‐based assay for detecting mpox virus. Mpox specific conserved sequences were identified in polA (E9L) gene which differ by a single nucleotide polymorphism (SNP) from all the viruses present in the genus Orthopoxvirus. This SNP was exploited in our assay to specifically distinguish mpox virus from other related orthopox viruses with a limit of detection of 1 copy/μl in 30 min. The assay exhibits a sensitive and specific detection of mpox virus which can prove to be of practical value for its surveillance in areas infected with multiple orthopox viruses, especially in hotspots of mpox virus infections.

Publisher

Wiley

Subject

Infectious Diseases,Virology

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