Deafness is a common sensory disorder, 70% of which are non-syndromic deafness (NSHL). There are currently more than 70 cloned genes associated with NSHL. This study used SNaPshot technology to explore the four hot spot mutations of deafness genes in 150 moderate-to-sense non-syndromic hereditary neurotic deafness cases in Yunnan, and to elucidate the epidemic distribution of deafness genes in ethnic minority NSHL population in Yunnan Province. This study enrolled 150 cases of Bai, Hani, and Yi non-syndromic deaf patients and 50 cases of Han NSHL patients as control in the special education school and the outpatient clinic of Yunnan province from January 2015 to January 2017. Medical history collection, hearing and imaging examinations, and 21 hot spot mutations of 4 deafness genes detection were performed. Molecular epidemiological control study was adopted. All patients underwent an otolaryngology specialist examination to diagnose the severity of sensorineural hearing loss. The deafness associated with the screening site in the control group was 36.0% (18/50), while it was 20.7% (31/150) in the experimental group. The mutation rates of three deafness-related genes, including GJB2, SLC26A4, and 12SrRNA in 150 ethnic minority deaf populations were 51.6% (16/31), 29% (9/31), and 19.4% (6/31), respectively. However, the GJB3 gene mutation was not detected. Mitochondria! 12SrRNA, GJB2-235delC, and SLC26A4-IVS7-2A > G were hotspot gene mutation.