A method for simultaneous detection of wild and mutant types of a virus

Abstract

A method is disclosed for the simultaneous detection of wild-type and Q57H mutant SARS-CoV-2 in a single assay. The method comprises reverse transcription of viral RNA using random hexamers, followed by multiplex PCR amplification using a combination of four specific primers targeting both conserved and mutation-specific regions. The amplified DNA products are resolved on a 2% agarose gel, yielding distinct band sizes corresponding to total SARS-CoV-2 presence (391 bp), wild-type (264 bp), and mutant (180 bp) variants. This method enables simple, rapid, and cost-effective identification of SARS-CoV-2 and its Q57H mutation in a single diagnostic workflow.