Helicobacter pylori which is the major agent causing peptic ulcer, gastric cancer and gastric lymphoma, is identified as a class I carcinogen. Invasive and non-invasive tests have been used in the diagnosis of H. pylori infection. Clarithromycin resistance in H. pylori strains is increasing nowadays, thus leading to failures in eradication therapy. Fluorescence in situ hybridization (FISH) method which can simultaneously detect the presence of H. pylori and clarithromycin resistance is a non-culture dependent molecular technique. In this review article the prevalence and clinical outcome of antibiotic resistance in H. pylori, molecular mechanisms of the resistance and molecular methods to detect the resistance, with priority of FISH technique have been discussed.