Fluorescent-Magnetic-Catalytic Nanospheres for Dual-Modality Detection of H9N2 Avian Influenza Virus

The outbreak of H9N2 avian influenza virus (H9N2 AIV) brings high mortality and huge economic losses every year. Sensitive and reliable detection methods are essential to timely diagnosis and treatment. Herein, a dual-modality immunoassay is proposed for H9N2 AIV detection by employing fluorescent-magnetic-catalytic nanospheres (FMCNs) as labels and alkaline phosphatase (ALP)-induced metallization as a signal amplification strategy. The excellent magnetic properties of FMCNs endow the assay a potential application in complex samples. Also, the excellent fluorescence properties of FMCNs enable fluorescence modality readout. The antibodies on the FMCN surface can achieve efficient capture and separation of targets. Amplified electrochemical modality readout can be obtained through ALP-catalyzed silver deposition. Dual-modality immunoassay combined the advantages of electrochemical assay with fluorescence assay and provides accurate detection results to meet different testing needs. With two quantitative analysis forms, H9N2 AIV can be detected by electrochemical signals with a quantitation range of 0.1 to 1000 ng/mL and a detection limit of 10 pg/mL. The linear range is 300 to 1000 ng/mL with a detection limit of 69.8 ng/mL by the fluorescence signal readout. Moreover, the specificity, anti-interference ability, accuracy, and diversity of the proposal have unlimited potential for early diagnosis of suspect infections.