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Abstract In the present investigation, mechanisms of carbapenems resistance in Ps. aeruginosa were studied. The studies were carried out on (53) carbapenems resistant Ps. aeruginosa isolates collected from Mansoura University hospitals. Sensitivity test was performed and MIC of imipenem and meropenem were determined. The study detected isolates which produced serine carbapenemases, metalloenzymes and AmpC β-lactamases. The biological activities of extracted β-lactamases against imipenem were determined. Studying characteristics of some β-lactamases by using spectrophotometric methods was performed through determination of their unit activities for both benzyl penicillin and imipenem and determination of rates of hydrolysis of benzyl penicillin, imipenem and meropenem. Also the effect of some inhibitors such as EDTA and clavulanic acid was studied and kinetic parameters of purified enzymes were determined. Separation and analysis of outer membrane protein by SDS-PAGE was performed. Genes of (intI-1, blaVIM and blaIMP) on plasmid DNA and (oprD, mexB and ampC) on chromosomal DNA were detected by PCR. Nucleotide sequencing of genes (blaVIM, oprD, and ampC) was studied by using Sanger’s technique. |