BACKGROUND: Recent antimicrobial resistance data for Neisseria gonorrhoeae are lacking in Uganda, where, until 2010, ciprofloxacin was the nationally recommended first-line treatment of presumptive gonorrhea. This study assessed the antimicrobial susceptibility patterns of N. gonorrhoeae isolates cultured from female sex workers (FSWs) in Kampala. METHODS: Gonococci were isolated from endocervical specimens collected from women enrolled in a FSW cohort for 18 months (2008-2009). Minimum inhibitory concentrations for 7 antibiotics (ciprofloxacin, cefixime, ceftriaxone, azithromycin, spectinomycin, penicillin, and tetracycline) were determined for 148 isolates using Etest strips. The European Committee on Antimicrobial Susceptibility Testing version 1.3 clinical breakpoints were used to assign susceptibility categories. The 2008 World Health Organization N. gonorrhoeae panel was used for quality assurance purposes. RESULTS: For ciprofloxacin, 123 (83.1%) gonococcal isolates were resistant, 2 (1.4%) had intermediate susceptibility, and 23 (15.6%) were fully susceptible. All isolates were susceptible to ceftriaxone and spectinomycin, whereas 1 isolate (0.7%) was resistant to cefixime. For azithromycin, 124 isolates (83.8%) were susceptible, 20 (13.5%) had decreased susceptibility, and 4 (2.7%) were resistant. Most isolates were resistant to penicillin (101; 68.2%) and tetracycline (144; 97.3%). The minimum inhibitory concentration ranges for each antibiotic were as follows: ciprofloxacin (0.002-32 mg/L), ceftriaxone (≤0.002-0.064 mg/L), cefixime (≤0.016-0.38 mg/L), spectinomycin (2-24 mg/L), azithromycin (0.023-1 mg/L), penicillin (0.094-32 mg/L), and tetracycline (0.019-256 mg/L). CONCLUSIONS: The high prevalence of ciprofloxacin-resistant gonorrhea observed in Kampala-based FSW emphasizes the need for sustainable gonococcal antimicrobial resistance surveillance programs in Uganda and, in general, Africa.