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Δευτέρα 10 Σεπτεμβρίου 2018

Expression of Pseudomonas aeruginosa antibiotic resistance genes varies greatly during infections in cystic fibrosis patients [Mechanisms of Resistance]

The lungs of individuals with cystic fibrosis (CF) become chronically infected with Pseudomonas aeruginosa that is difficult to eradicate by antibiotic treatment. Two key P. aeruginosa antibiotic resistance mechanisms are the AmpC β-lactamase that degrades β-lactam antibiotics and MexXYOprM, a three-protein efflux pump that expels aminoglycoside antibiotics from the bacterial cells. Levels of antibiotic resistance gene expression are likely to be a key factor in antibiotic resistance but have not been determined during infection. The aims of this research were to investigate expression of the ampC and mexX genes during infection in patients with CF and in bacteria isolated from the same patients. P. aeruginosa isolates from 36 CF patients were grown in laboratory culture and gene expression measured by reverse transcription quantitative PCR (RT-qPCR). Expression of ampC varied over 20,000-fold and that of mexX over 2000-fold between isolates. Expression levels of both genes were on average increased by the presence of sub-inhibitory concentrations of antibiotics. To measure P. aeruginosa gene expression during infection, we carried out RT-qPCR using RNA extracted from fresh sputum samples obtained from 31 patients. Expression of ampC varied over 4000-fold, while mexX expression varied over 100-fold, between patients. Despite these wide variations, median levels of expression of ampC in sputum were similar to those in laboratory-grown bacteria. Expression of mexX was higher in sputum than in laboratory-grown bacteria. Overall, our data demonstrate that genes that contribute to antibiotic resistance can be highly expressed in patients but there is extensive isolate-to-isolate and patient-to-patient variation.



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