Journal
SENSORS AND ACTUATORS B-CHEMICAL
Volume 282, Issue -, Pages 945-951Publisher
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2018.11.147
Keywords
Pathogen detection; Isothermal amplification; Deoxyribozyme cascade; G-quadruplex peroxidase; Colorimetric test; NASBA
Funding
- Florida Department of Health, Biomedical Research Program [7ZK33]
- National Institute of Allergy and Infectious Diseases of the National Institutes of Health [R21AI123876, R01AI110692]
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A colorimetric nucleic acid based test for label-free pathogen detection has been developed and used for the detection of the Zika virus. The test relies on nucleic acid sequence-based amplification (NASBA) of a viral RNA followed by interrogation of the amplicon by a cascade of deoxyribozymes constituting a visual split deoxyribozyme (vsDz) probe. The probe consists of a split phosphodiesterase deoxyribozyme, which forms its catalytic core upon binding to a specific amplicon fragment. The catalytically active complex recognizes and cleaves an inhibited peroxidase-like deoxyribozyme (PDz), thereby activating it. Active PDz catalyzes hydrogen peroxide-mediated oxidation of a colorless substrate into a colored product, thereby generating a visible signal. Viral RNA (10(6) copies/mL or higher) triggers intense color within 2 h. The test selectively differentiates between Zika and closely related dengue and West Nile viruses. The reported technology combines isothermal amplification and visual detection and therefore represents a basis for the future development of a cost-efficient and instrument-free method for point-of-care nucleic acid analysis.
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