Abstract: The present invention relates to the field of virology, and specifically discloses a short peptide having a dengue virus replication inhibition function and an application thereof. The amino acid sequence of the short peptide provided in the present invention is KHGHHRH, i.e. Lys-His-Gly-His-His-Arg-His (SEQ ID NO. 1). The short peptide has a high specificity affinity with NS5 and has the function of efficiently inhibiting dengue virus replication, the anti-viral effect thereof not been limited to DENV-2, but also having a significant inhibitory effect on the replication of type 1, type 3, and type 4 dengue virus. One cysteine is added to the two ends of the short peptide sequence, the short peptide being cyclised by means of the cysteines at the two ends to form a cyclic peptide. The obtained cyclic peptide strengthens the dengue virus replication inhibition function, and can be used for specific treatment of dengue virus infection.