A method of identifying a Mycobacterium using surface enhanced Raman spectroscopy (SERS), disclosed herein, comprises disposing a sample suspected to comprise a Mycobacterium on a SERS-active substrate, detecting surface enhanced Raman signals corresponding to an alpha-mycolic acid, a methoxy-mycolic acid, and a keto-mycolic acid from the sample disposed on the SERS-active substrate, and determining one or more ratios of intensity of the surface enhanced Raman signals to identify the Mycobacterium. Disclosed herein also includes a method of detecting a mycobacterial disease using surface enhanced Raman spectroscopy (SERS), the method comprising identifying a Mycobacterium according to the method described above, and determining the mycobacterial disease based on the Mycobacterium identified.