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Background  Contamination of endoscopy equipment by  Helicobacter pylori  ( H. pylori ) frequently occurs after endoscopic examination of  H. pylori -infected patients. In the hospital, manual pre-cleaning and soaking in glutaraldehyde is an important process to disinfect endoscopes. However, this might not be sufficient to remove  H. pylori  completely, and some glutaraldehyde-resistant bacteria might survive and be passed to the next patient undergoing endoscopic examination through unidentified mechanisms. We identified an Imp/OstA protein associated with glutaraldehyde resistance in a clinical strain, NTUH-C1, from our previous study. To better understand and manage the problem of glutaraldehyde resistance, we further investigated its mechanism.    Results  The minimal inhibitory concentrations (MICs) of glutaraldehyde andexpression of  imp/ostA  RNA in 11 clinical isolates from the National Taiwan University Hospital were determined. After glutaraldehyde treatment, RNA expression in the strains with the MICs of 4–10 μg/ml was higher than that in strains with the MICs of 1–3 μg/ml. We examined the full-genome expression of strain NTUH-S1 after glutaraldehyde treatment using a microarray and found that 40 genes were upregulated and 31 genes were downregulated. Among the upregulated genes,  imp/ostA  and  msbA , two putative lipopolysaccharide biogenesis genes, were selected for further characterization. The sensitivity to glutaraldehyde or hydrophobic drugs increased in both of  imp/ostA  and  msbA  single mutants. The  imp/ostA  and  msbA  double mutant was also hypersensitive to these chemicals. The lipopolysaccharide contents decreased in individual  imp/ostA  and  msbA  mutants and dramatically reduced in the  imp/ostA  and  msbA  double mutant. Outer membrane permeability assay demonstrated that the  imp/ostA  and  msbA  double mutation resulted in the increase of outer membrane permeability. Ethidium bromide accumulation assay demonstrated that MsbA was involved in efflux of hydrophobic drugs.    Conclusion  The expression levels of  imp/ostA  and  msbA  were correlated with glutaraldehyde resistance in clinical isolates after glutaraldehyde treatment. Imp/OstA and MsbA play a synergistic role in hydrophobic drugs resistance and lipopolysaccharide biogenesis in  H. pylori .

Synergistic effect of imp/ostA and msbA in hydrophobic drug resistance of Helicobacter pylori