AIMS: To evaluate a technique for culture of Helicobacter pylori in large quantities of liquid media and to determine the factors that could influence the results. METHODS: Fifteen clinical isolates of H pylori and a reference strain of H pylori NCTC11637 were used to evaluate a method to cultivate the organism in 100 ml liquid medium comprising brain heart infusion broth with 5% horse serum and 0.25% yeast extract. Tissue culture flasks containing the inoculated liquid medium were placed in a CO2 incubator with 5% CO2 for 2 hours and then incubated in a shaking incubator at 120 rpm. RESULTS: All the clinical isolates and the reference strain grew in the broth, although only a moderate growth of the reference strain occurred. Inoculum size significantly influenced the kinetics of growth of H pylori in the liquid medium. Vancomycin, nalidixic acid, and amphotericin B, used to suppress contamination, did not affect growth of H pylori in the medium. CO2 was essential for H pylori to grow or survive in the liquid medium. Incubation with CO2 in a CO2 incubator for 30 minutes or 2 hours did not affect the results. CONCLUSIONS: H pylori can be cultivated in large quantities of brain heart infusion broth with 5% horse serum and 0.25% yeast extract. Initial inoculum concentrations influence the kinetics of H pylori growth in the liquid medium. Vancomycin, nalidixic acid, and amphotericin B can be used as selective antimicrobial agents. CO2 is essential for initial growth of H pylori in liquid media. The findings in this study may provide a useful, reproducible, and simple method for biochemical, molecular, and physiological studies of H pylori, when those require large quantities of the organism.
AIMS: To evaluate a technique for culture of Helicobacter pylori in large quantities of liquid media and to determine the factors that could influence the results. METHODS: Fifteen clinical isolates of H pylori and a reference strain of H pylori NCTC11637 were used to evaluate a method to cultivate the organism in 100 ml liquid medium comprising brain heart infusion broth with 5% horse serum and 0.25% yeast extract. Tissue culture flasks containing the inoculated liquid medium were placed in a CO2 incubator with 5% CO2 for 2 hours and then incubated in a shaking incubator at 120 rpm. RESULTS: All the clinical isolates and the reference strain grew in the broth, although only a moderate growth of the reference strain occurred. Inoculum size significantly influenced the kinetics of growth of H pylori in the liquid medium. Vancomycin, nalidixic acid, and amphotericin B, used to suppress contamination, did not affect growth of H pylori in the medium. CO2 was essential for H pylori to grow or survive in the liquid medium. Incubation with CO2 in a CO2 incubator for 30 minutes or 2 hours did not affect the results. CONCLUSIONS: H pylori can be cultivated in large quantities of brain heart infusion broth with 5% horse serum and 0.25% yeast extract. Initial inoculum concentrations influence the kinetics of H pylori growth in the liquid medium. Vancomycin, nalidixic acid, and amphotericin B can be used as selective antimicrobial agents. CO2 is essential for initial growth of H pylori in liquid media. The findings in this study may provide a useful, reproducible, and simple method for biochemical, molecular, and physiological studies of H pylori, when those require large quantities of the organism.
Authors: S Krajden; J Bohnen; J Anderson; J Kempston; M Fuksa; A Matlow; N Marcon; G Haber; P Kortan; M Karmali Journal: J Clin Microbiol Date: 1987-06 Impact factor: 5.948
Authors: Harry Hua-Xiang Xia; Shiu Kum Lam; Annie O O Chan; Marie Chia Mi Lin; Hsiang Fu Kung; Keiji Ogura; Douglas E Berg; Benjamin Chun-Yu Wong Journal: World J Gastroenterol Date: 2005-04-07 Impact factor: 5.742
Authors: L K Siu; W K Leung; A F Cheng; J Y Sung; T K Ling; J M Ling; E K Ng; J Y Lau; S C Chung Journal: J Clin Microbiol Date: 1998-10 Impact factor: 5.948
Authors: Alba E Vega; Teresa I Cortiñas; Claudia M Mattana; Humberto J Silva; Olga Puig De Centorbi Journal: J Clin Microbiol Date: 2003-12 Impact factor: 5.948