A Novel Assay for Easy and Rapid Quantification of Helicobacter pylori Adhesion

作者：Skindersoe Mette E^*; Rasmussen Lone; Andersen Leif P; Krogfelt Karen A

来源：Helicobacter, 2015, 20(3): 199-205.

DOI：10.1111/hel.12191

• Helicobacter pylori
• sabA
• babA
• adhesion
• alamarBlue
• resazurin

BackgroundReducing adhesion of Helicobacter pylori to gastric epithelial cells could be a new way to counteract infections with this organism. We here present a novel method for quantification of Helicobacter pylori adhesion to cells.
MethodsHelicobacter pylori is allowed to adhere to AGS or MKN45g cells in a 96-well microtiter plate. Then wells are added saponin, which lyses the cells without affecting the bacteria. After addition of alamarBlue((R)) (resazurin) and 1- to 2-hour incubation, fluorescence measurements can be used to quantify the number of adherent bacteria.
ResultsBy use of the method, we demonstrate that adhesion of both a sabA and babA deletion mutant of H.pylori is significantly reduced compared to the wild type.
ConclusionThe method offers a number of applications and may be used to compare the adherence potential of different strains of H.pylori to either cells or different materials or to screen for potential anti-adhesive compounds. The results presented here suggest that this easy and reproducible assay is well suited for quantitative investigation of H.pylori adhesion.

• 出版日期2015-6