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Background and objectives: Neisseria gonorrhoeae is the second most sexually transmitted diseases agents in developing countries. Antimicrobial resistance strains have created serious health concern. The aim of this study was to determine the frequency of endocervical gonococcal infection and antimicrobial susceptibility of N.gonorrhoeae in Kashan, Iran. Materials and Methods: In this study, 294 endocervical swabs were collected from married women referred to the obstetrics and gynecology clinics in Kashan from December 2012 to May 2013. The samples were cultured in modified Thayer Martin in 37°C with 5-10% CO2 for 72 hours. Gram staining, oxidase, catalase and carbohydrate utilization tests were used to identify the isolated species. All isolates were tested for their susceptibilities to antimicrobials using the Kirby Bauer-disk diffusion techniques. Results: N.gonorrhoeae was detected in 2.38% of studied cases (95% confidence interval [CI] 1.5-3.26%). All isolates were resistance to ceftriaxone, penicillin G, ciprofloxacin, cefepime, and two isolate (28.5%) showed intermediate sensitivity to tetracycline. Conclusion: Continued monitoring of prevalence of N.gonorrhoeae is important for preventing the dissemination of this microorganism. The present study emphasizes the importance of surveillance of antimicrobial resistance of N.gonorrhoeae in order to manage the rate of resistant strains and to revise the treatment policies.