Abstract: Influenza A viruses (IAVs) exhibit rapid mutation and high infectivity, leading to a considerable number of cases and deaths globally. Thus, the development of broad-spectrum inhibitors is of paramount significance. In this study, targeting the hemagglutinin (HA) of IAVs, a set of high-affinity and specific aptamers, namely Apt(HA)-1, Apt(HA)-4, Apt(HA)-9, Apt(HA)-25, and Apt(HA)-93520, were obtained through CE-SELEX. The dissociation constants (K_D) of this group of aptamers Apts(HA) with the HA protein were determined by the SPR method to be 25.9, 44.8, 50.7, 54.0, and 45.2 nmol·L^-1, respectively. Apts(HA) can recognize and detect the internationally common influenza virus strains A/PR/8/34 (H1N1) and A/FM/1/47 (H1N1), as well as the clinically isolated Tamiflu-resistant strain A/Liaoningzhenxing/1109/2010 (H1N1), amantadine-resistant strains A/Hunanzhuhui/1222/2010 (H3N2) and A/Fujiantongan/196/2009 (H3N2), and possess broad-spectrum inhibitory effects on influenza viruses. These aptamer inhibitors feature strong in vitro accessibility, good stability, simple structure, and small size. This study not only developed novel broad-spectrum aptamers against IAVs, exerting extensive inhibitory effects on various IAVs, but also paved the way for an effective strategy in the development of universal virus inhibitors.