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Volume: 5, Issue: 5, Sep-Oct, 2017
DOI: 10.7324/JABB.2017.50501

Research Article

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


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

  Author Affiliations


Abstract

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.

Keywords:

Duplex specific nuclease; norovirus; viral detection; RNA; RNA detection; environmental water.



Citation: Wurtzler EM, Ravi R, Kapoor V, Wendell D. Rational Design of Duplex Specific Nuclease for One-Step Isothermal Viral RNA Detection. J App Biol Biotech. 2017; 5 (05): 1-10.


Copyright: Author(s). This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License, which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited.

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