Premium
Sequential Statistical Improvement of the Liquid Cultivation of Helicobacter pylori
Author(s) -
DuqueJamaica Rocio,
ArévaloGalvis Azucena,
PoutouPiñales Raúl A.,
Trespalacios Alba A.
Publication year - 2010
Publication title -
helicobacter
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.206
H-Index - 79
eISSN - 1523-5378
pISSN - 1083-4389
DOI - 10.1111/j.1523-5378.2010.00763.x
Subject(s) - box–behnken design , helicobacter pylori , factorial experiment , agarose , chromatography , food science , chemistry , mathematics , statistics , medicine , response surface methodology
Background: Colonization of the gastric mucosa by Helicobacter pylori is one of the most important causes of acute and chronic gastric pathologies in humans. Achieving the growth of H. pylori in liquid media is of great importance in the development of clinical studies. In this study, we developed a sequential optimization strategy based on statistical models to improve the conditions of liquid culture of H. pylori . Materials and Methods: Four statistical models were sequentially used. First, a Box‐Behnken design was used to select the best process conditions (shaking speed, inoculum concentration, and final volume of culture). Secondly, a general factorial design was used to evaluate the influence of adding gel blocks or gel beads (shape and composition). Then a D‐optimal reduce design was carried out to allow the selection of the most influential factors in increasing the cell concentration (culture media components). Finally, another Box‐Behnken design was used to optimize the concentration of the culture media components previously selected. Results: After 12 hours of liquid culture a concentration of 25 × 10 8 cells per mL (9.4 log 10 cells per mL) of H. pylori was obtained, compared with a predicted 32 × 10 8 (9.5 log 10 cells per mL), which means between 1 and 5 log 10 units higher than some previous reports. Conclusions: The sequential statistical approach increased the planktonic H. pylori cell culture. The final culture media and conditions were: Brain Heart Infusion, blood agarose (1.5% w/v), lamb’s blood (3.18% v/v), DENT (0.11% v/v), and Vitox (0.52% v/v) at 60 rpm and 37 °C with filtered CO 2 (5% v/v) bubbled directly into the culture media in a final volume of 76.22 mL.