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Rational Design of Duplex Specific Nuclease for One-Step Isothermal Viral RNA Detection

Elizabeth M. Wurtzler, Ranjani Ravi, Vikram Kapoor, David Wendell.

RNA viruses are a potent human adversary, evidenced by several global pandemics including the Ebolavirus in West Africa, the emerging Zika virus, and outbreaks of new Influenza strains and Norwalk virus in the food supply and cruise ships. Despite the virulence of these pathogens, there remains a significant limitation for detecting these viruses in a fast, accurate and cost effective manner. To meet this need we present a modified form of the duplex specific nuclease enzyme from the Paralithodes camtschaticus crab capable of generating an RNA-based signal amplification in a fraction of the time required for standard RT-qPCR. The applicability of this enzyme is demonstrated in an assay for Norwalk virus detection with a lower limit of ~100 viral copies per liter of environmental water.

Key words: Duplex Specific Nuclease; Norovirus; viral detection; RNA; RNA detection; environmental water

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Journal of Contemporary Medical Education


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