Dent s supplement

A total of 88 non-repetitive H. pylori clinical isolates were obtained from patients (47 females and 41 males) recruited in previous studies (research no. ETP-2013-042 and GUP-2011-307) between 2011–2015. Patients (15 Malays, 52 Chinese and 21 Indians) had a mean age of 54.68 ± 16.87 years ranging from 17 to 83 years. In these projects, gastric biopsies were taken from the antrum and/or corpus of the patients’ stomach for H. pylori culture and used as one of the diagnotic methods for determination of the H. pylori infection status in patients. H. pylori isolates grew from the culture method during the studies were stored at −70 °C in Brucella broth containing 15% glycerol. Histopathological examination was performed on gastric biopsies in both studies to detect the presence of H. pylori along with grading for gastritis. H. pylori were subcultured from frozen stock onto Columbia blood agar (Oxoid, Basingstoke, England) supplemented with 7% sheep blood and Dent’s supplement (Oxoid, Basingstoke, England) and incubated at 37 °C for five to seven days under microaerophilic environment.

Biopsies for culture were placed in Brucella broth (BBL; Becton Dickson, USA) containing 15% glycerol and transported to the Microbiology laboratory in UKMMC. Gastric biopsies were plated onto Columbia agar (Oxoid, Basingstoke, UK) supplemented with 7% defibrinated sheep blood (Oxoid) and Dent’s supplement (Oxoid). The inoculated agar plates were incubated at 37°C up to 5–10 days in microaerophilic conditions generated using microaerophilic CampyGen gas pack (Oxoid). Culture was recorded as positive when H. pylori growth identified by Gram staining showed gram-negative curved/spiral rods and positive for urease, catalase and oxidase tests.20 (link) Colonies were subcultured on several agar plates and used for antibiotic susceptibility testing and DNA extraction.