Abstract: Objective To investigate the incidence of common deafness gene mutations in patients with non-syndromic deafness. Methods The genomic DNA of 240 deafness patients were extracted from their peripheral blood. The hot-spot deafness gene mutations, including GJB2, GJB3, SLC26A4 and mitochondrial 12S rRNA genes, were detected using a nine deafness gene mutations detection kit. Results Various types of gene locus mutations were detected in 102 of the 240 patients, including 44 GJB2 gene mutations (18.33%, 44/240), 38 SLC26A4 gene mutations (15.83%, 38/240), 17 mitochondrial 12S rRNA gene mutations (7.08%, 17/240). The diagnosis results suggested that 60 cases were sure to be hereditary hearing loss and 42 cases were suspected hereditary hearing loss, which accounted for 42.5% (102/240) of the total. Conclusion Non-syndromic deafness patients have a high ratio of carrying deafness gene, and molecular genetic screening for these hot-spot mutations is a pivotal step for the prevention and control of genetic deafness.