The genus Gordonia is one of the mycolic acid-containing aerobic actinomycetes. This genus has 38 named species that are widespread in the natural environment; however, Gordonia species rarely cause human infections. A 76-year-old woman presented with cough and sputum for over 1 year and was suspected of having nontuberculous mycobacterial (NTM) lung disease. An NTM isolate from the sputum was initially identified as Mycobacterium lentiflavum or Mycobacterium genavense by genotypic identification targeting internal transcribed spacer (ITS). However, the isolate was finally confirmed as Gordonia otitidis by sequencing of 16S rRNA, gyrB and secA1 genes. In patients with suspected NTM lung disease, the etiologic agent might be an organism other than NTM such as G. otitidis but still be identified as NTM without sequencing of 16S rRNA or other genes. Especially in case that a possible NTM isolate is identified as M. lentiflavum or M. genavense by the genotypic method targeting ITS, additional genotypic tests such as sequencing of 16S rRNA and other genes would be necessary for more reliable identification.