Fluoroimmunoassay of influenza virus using sulfur-doped graphitic carbon nitride quantum dots coupled with Ag

Novel sulfur-doped graphitic carbon nitride quantum dots (S-gCNQDs) are synthesized using a single-source precursor in a one-step solvothermal process. The S-gCNQDs with a size of ~ 5-nm displayed a strong green intrinsic fluorescence at 512 nm when excited at 400 nm, with a quantum yield of ~ 33% in aqueous solution. The prepared S-gCNQDs and Ag2S nanocrystals were applied as innovative functional materials to fabricate a biosensor for virus detection based on the conjugation of specific anti-human influenza A monoclonal antibody to the S-gCNQDs and Ag2S NCs, respectively. In the presence of the influenza A virus, an interaction between the S-gCNQDs/Ag2S-labeled antibody resulted in the formation of a nanosandwich structure, which is accompanied by the fluorescence enhancement of the S-gCNQDs. The change in fluorescence intensity linearly correlats with the concentration of the influenza A virus (H1N1) in the 10 fg/mL to 1.0 ng/mL range, with a limit of detection of 5.5 fg/mL. The assay was applied to the assay of clinically isolated influenza A virus (H3N2/Yokohama) mixed with human serum. The obtained limit of detection was 100 PFU/mL within the detection range of 102- 5 × 104 PFU/mL for the H3N2 virus. Graphical abstract.