Introduction Helicobacter pylori is usually acquired in early childhood. The systematic use of standard eradication therapy regimes has resulted in a rising prevalence of antibiotic-resistant strains and a decreasing efficiency of H. pylori eradication therapy. The latest ESPGHAN guidelines (2016) suggest only investigating H. pylori in paediatric patients who would benefit from treatments and to base the eradication therapy on susceptibility testing. Our local practice has been to test symptomatic children with H. pylori stool antigen and to treat with a standard triple therapy as a first line. If a second attempt at eradication fails, children have oesophagogastroduodenoscopy (OGD) on a dedicated gastroenterology/microbiology H. pylori culture list. Subsequent antibiotic therapy is then based on antibiotic sensitivity. Aims Our primary objective was to study the benefit of H. pylori eradication therapy based on culture and sensitivity in our population. Our secondary objective was to describe the sensitivities of the enumerated H. pylori. Subjects and Methods We retrospectively included all paediatric patients who had undergone OGD for H. pylori culture in Royal Hospital for Children, Glasgow, between 2014 and 2020. We collected data from patient electronic records. H. pylori colonisation was based on the presence/absence of the organism on histopathology. Eradication was assessed by either H. pylori stool antigen or subsequent gastric biopsy histopathology. Results In total, 20 patients were included with a median age of 10.1 [7.6–12.4] years. In keeping with our local practice they had a median of 2 attempts at eradication therapy before being referred for H. pylori culture. On these 20 patients, 15 patients (75%) had a confirmed colonisation by H. pylori on histopathology. 14 of these patients (93%) had successful culture of H. pylori, 1 (7%) had a failed culture. On these 14 cultures, 1(7%) had initial growth but failed sensitivities. 3 H. pylori cultures (21% of positive culture) were fully sensitive to amoxicillin, metronidazole and clarithromycin. 4 (28%) were resistant to a single agent (50% to metronidazole and 50% to clarithromycin). 5 (35%) were resistant to both clarithromycin and another agent (80% to metronidazole and 20% to amoxicillin) and 1 (7%) was fully resistant to these 3 antibiotics. After sensitivity-based eradication therapy, 7 patients (50% of positive cultures) had ongoing H. pylori colonisation (3 confirmed on repeated OGD, 4 confirmed on a positive stool antigen), 4 (28%) had confirmed eradication (3 confirmed on repeat OGD and 1 on negative stool antigen), and 2 results are still awaited. Out of the 7 patients who failed eradication, 6 (85%) patients had resistant organisms and the remaining patient had poor treatment compliance. Conclusion Our dedicated H. pylori list has excellent culture recovery (93%), but its set-up can be challenging. Our approach to restricting culture to two therapy failures may explain our high rates of targeted antibiotic failure (50%), but these patients may also be biased towards therapeutic non-compliance. Having a better knowledge of the positivity rate of stool antigens in our paediatric population and eradication rate after first-line therapy would be helpful to consciously consider our results.