Objectives: The aim of the study is to describe in detail the methods of microbiological diagnosis of Neisseria gonorrhoeae and sampling methods for diagnosing gonorrhea, to indicate possible ways to prevent gonorrhea infections and to examine the frequency of testing for N. gonorrhoeae in Split-Dalmatia County and the frequency of positive results. Material and methods: In this study, the materials and data of the Teaching Institute for Public Health of the Split-Dalmatia County were used. Neisseria gonorrhoeae are gram-negative diplococci that do not form spores. A nucleic acid amplification test is used to diagnose gonorrhea. However, positive results must be confirmed by another method. The diagnosis can also be made by direct microscopy, most often from urethral or endocervical smears, stained by Methylene or Gram, where intracellular diplococci are seen in polymorphonuclear leukocytes. Also, gonococcal cultivation is performed from a sample and an antibiogram can be used to obtain gonococcal susceptibility data. Results: Through a five-year period from 1.1.2016. to 31.12.2020. a total of 136 samples tested by the method of aerobic cultivation for the bacterium N. gonorrhoeae were received at the Teaching Institute for Public Health of the Split-Dalmatia County. Of these, 127 were negative, while N. gonorrhoeae was detected in 9 (6.6%) samples, all of which were male. The PCR method on N. gonorrhoeae tested 187 samples, of which 172 were negative and N. gonorrhoeae was detected in 15 (8%) samples. There were 12 positive men (8.8%), 1 positive woman (4.2%), and 2 positive results in the first urine stream. Conclusion: Gonorrhea is a sexually transmitted disease that most often occurs as an asymptomatic disease, and when symptoms appear, it is a purulent discharge from the genitals, lower abdominal pain, pain during intercourse, postpartum hemorrhage, sterility, etc. An increasing problem is the development of reduced sensitivity to drugs where resistance to penicillins, tetracyclines and quinolones has been found. Both partners need to be treated because if they are not treated at the same time, the disease will reactivate.