Quinolone Resistance Detection by PCR-RFLP and Multiplex-PCR among Extended- Spectrum β- Lactamase Producing Enterobacteriaceae

Abstract

Quinolone resistance limits the therapeutic potential for the Extended-Spectrum β-lactamase (ESBL) producing Enterobacteriaceae. The aim of this study was to investigate the most common mechanisms of quinolones resistance in ESBL-producing Enterobacteriaceae using both phenotypic and genotypic methods. Out of 1766 clinical isolates collected between October 2012 and September 2013, 219 Enterobacteriaceae clinical isolates were ESBL producers, nalidixic acid and ciprofloxacin resistant were identified by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF/MS) and the Minimal Inhibitory Concentration (MIC) values of ciprofloxacin and nalidixic acid wasdetected before and after addition of phenylalanine-arginine β-naphthylamide (PaβN) efflux pump inhibitor. Thirty three isolates were selected for screening of the plasmid-mediated fluoroquinolone resistance (PMQR) genes; (qnr, aac(6')-Ib-cr and qepA) and the efflux pump genes (oqxAB genes) by multiplex PCR. Whereas GyrA and ParC genes mutations were detected by PCR-RFLP assay. The PaβN changed the MIC values of 28 isolates. The GyrA gene mutation was detected in 24/33 (72.7%), while the par C gene mutation was detected in 3/33 (9.1%). Qnr-genes were detected in 13/33 (39.4%), aac(6')-Ib gene was detected in 24/33 (72.7%). qepA gene was detected in only one Klebsiella pneumoniae isolate. Finally the oqxA gene was detected in 16/33 (48.5%) of the studied isolates. The present study indicated that the PaβN was an effective phenotypic screening method for quinolones resistance efflux pump; moreover PCR-RFLP offered simple and rapid method for detection of ciprofloxacin-resistance that could be useful for clinical diagnosis and epidemiological studies

<p>Detection of Chromosomal and Plasmid-Mediated Quinolone Resistance Among <em>Escherichia coli</em> Isolated from Urinary Tract Infection Cases; Zagazig University Hospitals, Egypt</p>

“…16 High prevalence of PMQR determinants was also reported by El-sayed Marei et al among both extended-spectrum beta-lactamases (ESBLs) and non-ESBLs producing Enterobacteriaceae isolates (93.1% and 76%, respectively) from UTI. 31 In another study carried out by Khalil et al, a prevalence of PMQR genes of 85.7% was found among ESBLs producing Enterobacteriaceae clinical isolates, with aac(6ʹ)-Ib-cr gene being the most prevalent (72.7%), followed by qnr genes (39.3%), 32 which comes consistent with our result.…”

Characterization of Integrons and Quinolone Resistance in Clinical Escherichia coli Isolates in Mansoura City, Egypt

“…PMQR was detected in 36/43 (83.72%) phenotypically fluoroquinolone-resistant isolates. Several studies showed similar results [ 39 , 40 ]. Other studies that investigated PMQR genes recorded variations in their prevalence rate even in the same country [ 41 – 44 ].…”

“…In previous studies, integron I was more frequent among Gram-negative bacteria [36][37][38]. [39,40]. Other studies that investigated PMQR genes recorded variations in their prevalence rate even in the same country [41][42][43][44].…”

Prevalence and mechanisms of ciprofloxacin resistance in Escherichia coli isolated from hospitalized patients, healthy carriers, and wastewaters in Iran

“…Similar to findings reported by Khalil, et.al. [ 55 ], we found that PaβN, as an efflux pump inhibitor, reduced the ciprofloxacin MIC in isolates containing OqxA/B supporting the importance of OqxA/B in ciprofloxacin resistance. It has been shown that the coexistence of PMQR determinates in E. coli could potentially cause higher levels of quinolone resistance [ 50 ].…”