title: A Novel Assay for Easy and Rapid Quantification of Helicobacter pylori Adhesion.
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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Microbiology and Infection Control, Statens Serum Institute, Artillerivej 5, DK-2300, Copenhagen S, Denmark.
authors.forename: Mette E
authors.initials: ME
authors.lastname: Skindersoe
authors.affiliation: Department of Clinical Microbiology 9321, Copenhagen University Hospital, Rigshospitalet, Blegdamsvej 9, DK-2100, Copenhagen, Denmark.|Department of Infection Control 9101, Copenhagen University Hospital, Rigshospitalet, Blegdamsvej 9, DK-2100, Copenhagen, Denmark.
authors.forename: Lone
authors.initials: L
authors.lastname: Rasmussen
authors.affiliation: Department of Clinical Microbiology 9321, Copenhagen University Hospital, Rigshospitalet, Blegdamsvej 9, DK-2100, Copenhagen, Denmark.|Department of Infection Control 9101, Copenhagen University Hospital, Rigshospitalet, Blegdamsvej 9, DK-2100, Copenhagen, Denmark.
authors.forename: Leif P
authors.initials: LP
authors.lastname: Andersen
authors.affiliation: Microbiology and Infection Control, Statens Serum Institute, Artillerivej 5, DK-2300, Copenhagen S, Denmark.
authors.forename: Karen A
authors.initials: KA
authors.lastname: Krogfelt
mesh_terms: D018829:Adhesins, Bacterial / Q000235:genetics*; D001422:Bacterial Adhesion; D045744:Cell Line, Tumor; D004847:Epithelial Cells / Q000382:microbiology; D016481:Helicobacter Infections / Q000382:microbiology*; D016480:Helicobacter pylori / Q000235:genetics / Q000502:physiology*; D006801:Humans; D007202:Indicators and Reagents; D009154:Mutation; D010078:Oxazines; D011485:Protein Binding; D013270:Stomach / Q000382:microbiology; D014966:Xanthenes
chemical_list: D018829:Adhesins, Bacterial; D007202:Indicators and Reagents; D010078:Oxazines; C500808:SabA protein, Helicobacter pylori; D014966:Xanthenes; C005843:resazurin
keywords: Helicobacter pylori; adhesion; alamarBlue; babA; resazurin; sabA