Comparative Genomics Revealed Multiple Helicobacter pylori Genes Associated with Biofilm Formation In Vitro

PLoS One. 2016 Nov 21;11(11):e0166835. doi: 10.1371/journal.pone.0166835. eCollection 2016.

Abstract

Background: Biofilm formation by Helicobacter pylori may be one of the factors influencing eradication outcome. However, genetic differences between good and poor biofilm forming strains have not been studied.

Materials and methods: Biofilm yield of 32 Helicobacter pylori strains (standard strain and 31 clinical strains) were determined by crystal-violet assay and grouped into poor, moderate and good biofilm forming groups. Whole genome sequencing of these 32 clinical strains was performed on the Illumina MiSeq platform. Annotation and comparison of the differences between the genomic sequences were carried out using RAST (Rapid Annotation using Subsystem Technology) and SEED viewer. Genes identified were confirmed using PCR.

Results: Genes identified to be associated with biofilm formation in H. pylori includes alpha (1,3)-fucosyltransferase, flagellar protein, 3 hypothetical proteins, outer membrane protein and a cag pathogenicity island protein. These genes play a role in bacterial motility, lipopolysaccharide (LPS) synthesis, Lewis antigen synthesis, adhesion and/or the type-IV secretion system (T4SS). Deletion of cagA and cagPAI confirmed that CagA and T4SS were involved in H. pylori biofilm formation.

Conclusions: Results from this study suggest that biofilm formation in H. pylori might be genetically determined and might be influenced by multiple genes. Good, moderate and poor biofilm forming strain might differ during the initiation of biofilm formation.

Publication types

  • Comparative Study

MeSH terms

  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism
  • Biofilms / growth & development*
  • Genome, Bacterial
  • Genomics / methods*
  • Helicobacter pylori / classification
  • Helicobacter pylori / genetics
  • Helicobacter pylori / physiology*
  • High-Throughput Nucleotide Sequencing
  • In Vitro Techniques
  • Sequence Analysis, DNA

Substances

  • Bacterial Proteins

Grants and funding

This study was funded in full by the University of Malaya-Ministry of Education (UM-MoE) High Impact Research (HIR) Grant UM.C/625/1/HIR/MoE/CHAN/13/3 (Account No. H-50001-A000030) and the National Medical Research Council (NMRC) Grant No. R-182-000-180-213. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.