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Monitoring the infection behavior of avian influenza viruses in vivo is crucial for understanding viral pathogenesis and preventing its epidemics among people.Recently,a number of viral labeling methods have been applied for tracking viral infection in vitro and in vivo,but most of them are unsatisfied,such as laborious or decreasing viral activity.Herein we explored an alternative bioorthogonal reaction,known as “click chemistry reaction,” for labeling live H5N1 pseudotype virus(H5N1p)in mice.