Background To investigate the current status and trend of Helicobacter pylori(H. pylori)antibiotics resistance over 5 years in Nanjing area.Then we evaluated the primary and post-treatment antibiotic resistance of H. pylori in Nanjing area. Methods During the period from July 2017 to December 2019, 1533 gastric mucosal specimens from patients with positive H. pylori confirmed by breath test or rapid urease test were collected for isolation and identify H. pylori. The agar dilution method was used for antibiotic resistance test. The antibiotic resistance of H. pylori were compared with that in the period from May 2014 to May 2015. Results The result showed that the resistance rates of H. pylori to amoxicillin, clarithromycin, levofloxacin, furazolidone, tetracycline and metronidazole were 2.74%, 47.03%, 33.59%, 0.91%, 0.52% and 80.76%, respectively in the period of July 2017 to December 2019. Compared with the period of May 2014 to May 2015, the resistance rate of metronidazole was the most obvious, from 60.5% to 80.76% in the period of July 2017 to December 2019, followed by clarithromycin from 29.5% to 47.03%, and levofloxacin from 14.7% to 33.59%. The resistance rates of H. pylori (primary Vs post-treatment) to amoxicillin, clarithromycin, levofloxacin, furazolidone, tetracycline and metronidazole were 1.83% Vs 6.08%, 38.62% Vs 77.81%, 27.41% Vs 56.23%, 0.58% Vs 2.13%, 0.33% Vs 1.22%, 78.57% Vs 88.75%, respectively during the period of July 2017 to December 2019. Conclusions Antibiotic resistance of H. pylori remained a problem for the effective eradication of this pathogen and its associated diseases in Nanjing area. For post-treatment eradication patients, clinicians should took into account regional antibiotic resistance rate, personal antibiotic exposure history, economic benefit ratio, adverse antibiotic reactions, antibiotic availability and other aspects.