An urgent need exists for a rapid, cost-effective, facile and reliable nucleic acid assays for mass screening to control and prevent the spread of emerging pandemic diseases such as COVID-19. This urgent need is not fully met by current diagnostic tools. We have developed a rapid and reliable nucleic acid assay for confirming the infection status of diseases and variants. The assay will use a mechanism of DNA-mediated proximity assembly circuit (DPAC), in which a DNA structural switch is used to trigger the co-assembly of an enzyme/cofactor system in response to the input of specific nucleic acid sequences (<b>Figure 1A&B</b>). The triggered enzyme reaction will produce a fluorescent or colorimetric signal for rapid detection. Based on DPAC, we further propose to develop a dNANOSIS (referred as DNA-NANO-Diagnosis) assay platform that integrates RNA extraction, isothermal amplification, and DPAC sensor (<b>Figure 1C</b>). The proposed dNANOSIS aims to rapidly detect viral RNA from collected samples within 20-60 mins. The proposed assay can be automated or be integrated into a user-friendly paper-based detection or lateral flow device for point-of-care use. The proposed development will have the value-added benefits of: (1) a sensitive and reliable assay for confirming virus infection status; (2) rapid results; (3) a simple operation supporting facile use; and (4) an affordable cost for disposable use. The DPAC sensor is a novel mechanism of engineering smart molecular devices that integrate molecular computation with specific target recognition and signal amplification on the nanoscale.

DNA-mediated proximity assembly circuits for rapid molecular sensing and point-of-care diagnosis

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ACS Fall 2022

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