Objectives: The aim of the study is to present the pecularities of laboratory diagnostics of samples in suspected Helicobacter infection in the period from 18.05.2020 – 31.12.2023 of the Department of Clinical Microbiology and Hospital Infections of the University Hospital Center Osijek. Study design: The study was conducted as a cross-sectional study. Subjects and methods: The study included patients whose samples (stool and/or biopsy of the gastric mucosa) were analyzed for suspected H. pylori infection. Diagnostics was based on two methods. One method was an antigen test that detects the presence of fecal antigen of bacteria, and the second method used was the cultivation of samples obtained by biopsy of the gastric mucosa. Results: 798 samples were received over three years. Of that, almost twice as many stool samples. There were significantly more positive biopsy samples in 2021 and 2022. There was no significant difference in the distribution of the sample according to gender and score, culture or AT. Stool samples were submitted by significantly younger subjects who had more negative stool findings. There was no age difference in positive findings. The sensitivity of Helicobacter was tested to amoxicyline, tetracycline, clarithromycin, metronidazole, levofloxacin and rifampicin. Over the years, bacteria have proven to be sensitive to amoxicyline, tetracycline, rifampicin and levofloxacin, while they were resistant to metronidazole and clarithromycin. Conclusion: The prevalence of helicobacter is high all over the world, while in Croatia the percentage is even higher. Thanks to ICT methods, it is easy to detect bacteria in the body. The ICT method is cheap, fast, easy to perform and highly specific. Also; antimicrobial susceptibility testing is extremely important to monitor the rise of resistance.