Abstract

The world’s tropical and sub-tropical regions are at risk for Zika and dengue virus (ZIKV, DENV) spread. Co-infection between ZIKV and DENV and repeated DENV infections of different serotypes increase the disease severity by impairing immune responses. Therefore, to avoid high risks of virus infection and achieve effective clinical management, development of diagnostic sensor for rapid and accurate identification of each virus is very important. Herein, we report a graphene oxide-based multiplexed detection strategy to make the differential diagnosis of Zika and dengue viruses by targeting virus nucleic acid. The fluorometric detection strategy is integrated with loop-mediated isothermal amplification assay to further increase sensitivity. In the presence of target virus, fluorescent PNA probes recognize the single-stranded loops of the cauliflower-like-shaped virus amplicon in a sequence-specific manner. Meanwhile, graphene oxide functions to distinguish the valid fluorescence signal as a selective fluorescence quencher for single-stranded PNA probes. The present strategy allows multiplexed detection of closely related viruses for specific discrimination of the virus nucleic acids with simplified molecular diagnostic steps, without requiring any nucleic acid isolation step.

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