From April 2006 to August 2007, a total of 146 Neisseria gonorrhoeae isolates collected from 139 male patients in Taipei, Taiwan, were analyzed by N. gonorrhoeae multiantigen sequence typing (NG-MAST) and antibiotic susceptibility testing. The resistance rates of all isolates to ciprofloxacin, cefpodoxime, and cefixime were 76.7 (112/146), 21.2 (31/146), and 16.4% (24/146), respectively. NG-MAST identified 71 sequence types (STs), of which 21 STs contained 2 to 21 isolates. The isolates that belonged to the three major ST clusters typically were from patients who had specific epidemiological characteristics (such as sexual orientation and human immunodeficiency virus status). The major ST clones exhibited distinct resistance profiles and are associated with specific groups at high risk of human immunodeficiency virus and syphilis infections.