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

Mette E. Skindersoe*, Lone Rasmussen, Leif P. Andersen, Karen A. Krogfelt

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

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.

Original languageEnglish
JournalHelicobacter
Volume20
Issue number3
Pages (from-to)199-205
Number of pages7
ISSN1083-4389
DOIs
Publication statusPublished - 1 Jun 2015
Externally publishedYes

Keywords

  • Adhesion
  • AlamarBlue
  • BabA
  • Helicobacter pylori
  • Resazurin
  • SabA

Cite this