Background The high mutation rate of influenza A virus hemagglutinin segment brings great challenges to its long-term effective testing and subtyping. Method We analyzed the conserved sequences of hemagglutinin subtype H1-H9 by breadth first, and designed primers for HA subtyping based on conserved sequences. Results Our conserved sequence searching method get high specificity conserved sequences on H1-H9 subtypes respectively. And PCR experiments show that primers based on conserved sequences can be used in influenza A virus HA subtyping. Conclusions Conserved sequences based primers are expected to be the long-term effective influenza A virus HA subtyping tools.