Helicobacter pylori (H. pylori) has been incriminated as the causative agent of most cases of gastric problems in human. So far, there is no one test agreed upon as a gold standard for the diagnosis of H. pylori infection. In the present study, 250 gastric biopsies were collected from male and female dyspeptic patients of different ages recruited to the Gastroenterology Unit atAhmed Maher Hospital, a big governmental hospital located at the west of Cairo metropolitan, the capital of Egypt, from November 2019 to December 2020. Samples were subjected to bacteriological studies to isolate and identify H. pylori. Fifty isolates were identified as Helicobacter spp. according to morphological and biochemical bases. The biochemicalbehavior was variable among different isolates. When a specific semi nested PCR assay was applied on DNA extracted from the helicobacter isolates, only 5 were identified as H. pylori while the rest was considered non-H. Pylori helicobacter where many of them have been isolated from human gastric samples in previous studies.Therefore, a reliable diagnosis is crucial for better eradication therapy of helicobacter-originated infections in human and animals. Results of more than one test are required to elucidate the real causative agents of human gastritis and to detect animal carriers of helicobacter.