PCR and microarray analysis of AmpC and ESBLs producing Pseudomonas aeruginosa isolates from intensive care units

Abstract

Detection of AmpC and ESBL producing P. aeruginosa by phenotypic methods is challenging, especially in low-income countries such as Pakistan. Therefore, a molecular method was developed for rapid detection of these resistance markers. A total of 303 clinical samples were collected from intensive care units (ICUs) of the Jinnah postgraduate medical centre (JPMC) Karachi, Pakistan. The isolates were identified by traditional and matrix-assisted laser desorption ionization–time-of-flight mass spectrometry (MALDI-TOF-MS). Isolates were phenotypically analyzed for AmpCs and ESBL by D-test and by double disc synergy, respectively. The Check MDR CT103 XL and PCR techniques were used for the detection AmpCs and ESBLs. Out of 303 isolates, 148 (48.8%) were P. aeruginosa. The resistance pattern of P. aeruginosa against piperacillin, cefatizidime and cefepime was 59.4%, 64.8% and 59.4% respectively. More than 60% isolates were resistant to aminoglycosides and ciprofloxacin. All (148) strains were found sensitive to colistin. Phenotypic ESBL prevalence was 8.8% whereas genotypic resistance was 29.1%. bla was the most prevalent ESBL. Although 25.67% of P. aeruginosa isolates were positive phenotypically for AmpC, microarray (Check-MDR) analysis did not detect chromosomally located AmpC in any of the isolates. VE

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