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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\u2010based assay for detecting mpox virus. Mpox specific conserved sequences were identified in <jats:italic>polA<\/jats:italic> (E9L) gene which differ by a single nucleotide polymorphism (SNP) from all the viruses present in the genus <jats:italic>Orthopoxvirus<\/jats:italic>. 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\/\u03bcl in 30 min. 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