Dual-mode, regenerated DNA motor for simultaneous detection of viral gene fragments and diagnosis of infectious disease

Abstract

This study presents a dual-mode and regenerated DNA motor powered by exonuclease III (Exo III) for the simultaneous detection of viral gene fragments. The detection methodology is categorized into two distinct operational modes. The first mode emphasizes the simultaneous detection of two viral gene fragments from a specific virus. The presence of both genes triggers the operation of the DNA motor, generating a singular signal output. This mode operates on an “AND” logical mechanism, which enhances the precision of positive case identification. The second mode facilitates the simultaneous detection of three viral gene fragments from three different viruses within a single assay. The presence of these genes activates their respective motors, yielding distinct signal outputs. This mode supports the multiplex detection of three target genes, thereby aiding in the identification of previously uncharacterized viruses infecting patients and alleviating the logistical and financial burdens associated with multiple testing procedures. The detection limit in the “AND” logical mode is at the aM level, while in the multiplex mode, it reaches the fM level, facilitating the sensitive detection of viral gene fragments. The DNA motor can be regenerated by separating and reconstituting the utilized orbits, enabling its reuse for up to seven cycles in the “AND” logical mode and five cycles in the multiplex mode. Accurate diagnoses were achieved for patients exhibiting upper respiratory symptoms. Therefore, the proposed motor offers a novel and regenerative approach for viral gene fragments detection, demonstrating significant promise for application in the clinical diagnosis of viral infectious diseases.