The prevalence of tick-borne viral diseases is increasing, representing a significant public health concern due to their substantial impact on human health. In recent years, two novel orthonairoviruses, Songling virus (SGLV) and Tamdy virus (TAMV), have been demonstrated to be associated with human febrile illnesses and tick epidemics. Moreover, Huangpi tick virus 1 (HpTV-1), a member of the same virus genus, has been identified in ticks from both the Shandong and Hubei provinces. In this study, we successfully isolated HpTV-1 for the first time from Haemaphysalis longicornis in Hubei Province. The genome morphology of this isolate is similar to that of orthonairovirus, and phylogenetic analysis indicates that it forms a distinct clade within SGLV and TAMV of the Nairoviridae family. In vitro and in vivo studies have demonstrated that HpTV-1 is capable of infecting both animal and human cell lines, and of causing organ lesions in the liver, spleen, and lungs in a mouse model. Furthermore, a serological survey identified antibodies against HpTV-1 in 35 serum samples from goats in Hubei Province, 13 of which contained neutralising antibodies with an inhibition rate of over 50%. This evidence suggests that HpTV-1 can infect goats. Our findings indicate that HpTV-1 may represent a potential pathogen with the capacity to impact animal and human health. Consequently, further studies and surveillance of HpTV-1 are recommended, with a particular focus on Hubei Province, as well as other relevant locations.