Aspergillus species are the most common causative agents involved in otomycosis. In this study, 45 Aspergillus isolates were obtained from patients with otomycosis in western China during 2013-2016. The aim of this study is to identify the Aspergillus isolates to the species level by using β-tubulin gene sequencing and to evaluate their in vitro susceptibility to nine antifungal drugs: amphotericin B, itraconazole, voriconazole, posaconazole, ravuconazole, isavuconazole, caspofungin, micafungin and anidulafungin according to CLSI M38-A2. Our results indicate that A. tubingensis (18/45) is the predominant Aspergillus species causing ear infections in western China, which is three times more than its sibling species A. niger (6/45) and A. welwitschiae (2/45). Other detected species were A. fumigatus (n = 8), A. terreus (n = 7) and A. flavus (n = 4). Antifungal susceptibility data indicate that triazoles and echinocandins are active against the most Aspergillus isolates. There are no significant differences in the susceptibility among A. niger, A. tubingensis and A. welwitschiae to each of the antifungals tested. One azole-resistant A. fumigatus isolate with a TR34/L98H mutation in the CYP51A gene and one posaconazole-resistant A. terreus isolate presented among the studied isolates. In conclusion, A. tubingensis is the most prevalent Aspergillus species causing otomycosis in western China. Posaconazole and echinocandins are potential drugs for treatment of otomycosis due to Aspergillus; however, in vivo efficacy remains to be determined.