Microbiology / Pharmacodynamics

In vitro pharmacoynamic evaluation of anti-infectious agents

  • Technics used

Antibiotic resistance assessments: MIC, checkerboard, Time-kill curves, Hollow fiber

MIC and checkerboard on fungi and mucorales

  • Equipment’s

40 m² BSL2 lab

3 PSMII, equipped for in vitro experiments

2 In  House Hollow Fiber setting up able to fit with model including 2 antibiotics with  different half-lives.

CO2 incubator for in vitro intracellular model of infections (Pseudomonas aeruginosa, Mycobacterium abscessus

Automated plating and counting of CFU

Molecular Biology

Characterization of resistance mechanisms

  • Quantification of gene expression: Resistance Gene expression analysis is fundamental to understand dynamic changes in a bacterium in the contact of antibiotic. We have developed the gene expression analysis by qRTPCR on many genes involved in the resistance development in various species as: Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae and Acinetobacter baumannii.
  • PCR and Sequencing: Amplification sequencing of DNA fragments allows to study the identification of mutations conferring for exemple resistance phenotypes during antibiotic exposure. We are regularly sequencing gene involvein the resistance of antibiotics to various species as Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae and Acinetobacter baumannii
  • Access to WGS/NGS platform

Genome engineering

  • Site-Directed Mutagenesis: Construction of bacterial gene-inactivating plasmids, Construction of mutants (Knock out, Knock In, Mutational insertion) in Enterobacteriaceae, A. Baumannii and aeruginosa even on MDR clinical isolates.
  • Construction of reporter gene: It allows to follow expression of a gene of interest by fluorescence or luminescence

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