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A Novel Assay for Easy and Rapid Quantification of Helicobacter pylori Adhesion
Mette E Skindersoe1, Lone Rasmussen2,3, Leif P Andersen2,3
1Microbiology and Infection Control, Statens Serum Institute, Artillerivej 5, DK-2300, Copenhagen S, Denmark.
Background:
Reducing 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.
Methods:
Helicobacter 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(®) (resazurin) and 1- to 2-hour incubation, fluorescence measurements can be used to quantify the number of adherent bacteria.
Results:
By 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.
Conclusion:
The 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.
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