Objective To explore the subtype distribution and the primary drug resistance (PDR) among human immunodeficiency virus-1(HIV-1) infected individuals in Hunan Province, and provide reference for the prevention and control as well as antiretroviral therapy (ART) of acquired immunodeficiency syndrome. Methods Demographical and epidemiological data of individuals newly diagnosed with HIV-1 infection and without receiving ART in a hospital's HIV clinic in Hunan Province from January 2021 to August 2022 were collected. Patients' serum was taken, ribonucleic acid was extracted, and two rounds of polymerase chain reaction for gene amplification was performed with the In-house method. PCR product was sequenced with Sanger method for analyzing clinical data, HIV-1 subtypes and drug resistance of patients. Results A total of 667 patients were recruited. 20 patients failed the genotypic drug resistance test, thus 647 newly diagnosed HIV-1 infected patients were studied. 53 HIV-1 infected patients had PDR, with a incidence of 8.19%. These patients were mainly male (88.68%), unmarried (71.70%), men who had sex with men (MSM, 60.38%), 21-40 years old group (58.49%), and local residents of Hunan Province (98.11%). A total of 15 subtypes of HIV-1 were detected, and the main subtype were CRF_01AE(35.70%), CRF07_BC (25.19%), B/C (18.86%) and CRF55_01B(7.88%). The differences of PDR incidence in patients with different HIV-1 subtypes were statistically significant (χ²=18.62, P=0.017). Drug resis-tance rates of protease inhibitors (PIs), nucleotide reverse transcriptase inhibitors (NRTIs), non-nucleotide reverse transcriptase inhibitors (NNRTIs) and integrase inhibitors (INSTIs) were 0.15%, 2.78%, 6.65%, and 0.31%, respectively. Conclusion The distribution of HIV-1 subtype in Hunan Province is complex, and the incidence of PDR has exceeded the low-level drug resistance warning line (< 5%) established by the World Health Organization. It is necessary to strengthen the prevention and control of the infection and block the transmission of drug-resistant virus strains.