Plasmid-Mediated Mechanism of Quinolone Resistance on E. coli Isolates from Different Clinical Samples

Document Type : Original Articles

Authors

1 College of Biotechnology, Al-Qasim Green University, Babylon Province, Iraq

2 Collage of Medicine, Microbiological Department, University of Babylon Province, Iraq

Abstract

Quinolone antimicrobials are widely used in clinical medicine due to their wide spectrum with high tissue penetration and ease of use; but increasing resistance with clinical use appears to be common in some bacterial pathogens, including Escherichia coli (E.coli). The aim of this study was to investigate plasmid-mediated quinolone resistance determinants (PMQR) including, qnrA, qnrB, and qnrS as the emerging mechanisms of quinolone resistance of E.coli isolates from different clinical sites in Karbala province, Iraq. A total of 200 clinical samples were collected from patients suffering from infections such as UTI, gastro enteritis (diarrhea), vaginitis, and wound infections; 30 samples were diagnosed as E.coli clinical strain from both sexes and different ages after identification by biochemical test, VITEK-2 compact system, and by molecular method using 16Sr DNA marker. Antimicrobial susceptibility and minimal inhibition concentration (MIC) testing for nalidixic acid, norfloxacin, ciprofloxacin, levofloxacin, and gatifloxacin was performed using the broth microdilution method. All strains were screened for PMQR genes qnrA, qnrB, and qnrS by the PCR method after DNA extraction from tested clinical isolates of E.coli. The results showed that E. coli is largely isolated from vaginal (40%) and urine (32%) samples, followed by wound infections (24%) and stools (21%).The high occurrence rate of E. coli(33.33%) isolates was observed in participants aged 31-45 years, while a lower occurrence (10%)was recorded in a group of ˃ 60-year-old female participants. Females have a notably increased frequency of E.coli compared to males, with the female to male ratio being 87%:13%. Molecular investigation showed the total percentage of E.coli isolates harboring qnr genes to be 21/30 (70%); this figure is composed of 14/30 isolates harboring qnr in combined or mixed form (46.66%) and 7/30 (23.33%) isolates harboring qnr in single form (3 isolates harboring qnrA alone, 1 isolate harboring qnrB alone, 3 isolates harboring qnrS alone).The prevalence rates of qnrA, qnrB, and qnrS were 40%, 43.33%, and 53.33%, respectively. The results also showed that among E.coli isolates encoding qnr genes A, B, and S, 24%, 12%, and 36% were resistant to nalidixic acid, respectively. Among those isolates carrying qnrA, qnrB, and qnrS genes, 15.8%, 5.3%, and 26.3%, respectively, were resistant to ciprofloxacin. Moreover, Norfloxacin resistance was seen in 20.0%, 5.0%, and 30.0% of E.coli isolates harboring qnr A, B, and S genes, respectively. Levofloxacin resistance was seen in 37.5%, 75.0%, and 37.5% of the isolates carrying the qnrA, qnrB, and qnrS genes, respectively. The lowest resistance rates of qnrA, B, and S-positive E.coli strains were against gatifloxacin (0,0, and 25%, respectively).A high prevalence of qnr genes enhances the increasing resistance rate of E.coli against the quinolone antibiotic under study.

Keywords

Main Subjects

Article Title [French]

Mécanisme a Médiation Plasmidique de la Résistance aux Quinolones sur les Isolats d'E. coli Provenant de Différents Echantillons Cliniques

Abstract [French]

Les quinolones antimicrobiennes sont largement utilisées en médecine clinique en raison de leur large spectre, de leur pénétration tissulaire élevée et de leur facilité d'utilisation; mais l'augmentation de la résistance avec l'utilisation clinique semble être courante chez certains agents pathogènes bactériens, y compris Escherichia coli (E. coli). Le but de cette étude était d'étudier les déterminants de la résistance aux quinolones à médiation plasmidique (RQMP), y compris qnrA, qnrB et qnrS en tant que mécanismes émergents de la résistance aux quinolones des isolats d'E. coli provenant de différents sites cliniques dans la province de Karbala, en Irak. Au total, 200 échantillons cliniques ont été collectés auprès de patients souffrant d'infections telles que les infections urinaires, la gastro-entérite (diarrhée), la vaginite et les infections des plaies; 30 échantillons ont été diagnostiqués comme étant une souche clinique d'E. coli des deux sexes et d'âges différents après identification par test biochimique, système compact VITEK-2 et par méthode moléculaire utilisant le marqueur ADN 16Sr. Les tests de sensibilité aux antimicrobiens et de concentration minimale d'inhibition (CMI) pour l'acide nalidixique, la norfloxacine, la ciprofloxacine, la lévofloxacine et la gatifloxacine ont été effectués à l'aide de la méthode de microdilution en bouillon. Toutes les souches ont été criblées pour les gènes PMQR qnrA, qnrB et qnrS par la méthode RCP après extraction de l'ADN à partir d'isolats cliniques testés d'E. coli. Les résultats ont montré qu'E. coli est largement isolé des échantillons vaginaux (40%) et d'urine (32%), suivi des infections des plaies (24%) et des selles (21%). Le taux d'occurrence élevé des isolats d'E. coli (33.33%) a été observé chez les participants âgés de 31 à 45 ans, tandis qu'un taux d'occurrence plus faible (10%) a été enregistré dans un groupe de participantes ˃ 60 ans. Les femelles ont une fréquence considérablement plus élevée d'E. coli par rapport aux mâles, le ratio femelle/mâle étant de 87%: 13%. L'enquête moléculaire a montré que le pourcentage total d'isolats d'E. coli contenant des gènes qnr était de 21/30 (70%); ce chiffre est composé de 14/30 isolats hébergeant qnr sous forme combinée ou mixte (46.66%) et 7/30 (23.33%) isolats hébergeant qnr sous forme unique (3 isolats hébergeant qnrA seul, 1 isolat hébergeant qnrB seul, 3 isolats hébergeant qnrS seul). Les taux de prévalence de qnrA, qnrB et qnrS étaient de 40%, 43.33% et 53.33%, respectivement. Les résultats ont également montré que parmi les isolats d'E. coli codant pour les gènes qnr A, B et S, 24%, 12% et 36% étaient respectivement résistants à l'acide nalidixique. Parmi les isolats portant les gènes qnrA, qnrB et qnrS, 15.8%, 5.3% et 26.3%, respectivement, étaient résistants à la ciprofloxacine. De plus, une résistance à la norfloxacine a été observée dans 20.0%, 5.0% et 30.0% des isolats d'E. coli hébergeant les gènes qnr A, B et S, respectivement. Une résistance à la lévofloxacine a été observée dans 37.5%, 75.0% et 37.5% des isolats portant les gènes qnrA, qnrB et qnrS, respectivement. Les taux de résistance les plus faibles des souches d'E. coli qnrA, B et S-positives étaient contre la gatifloxacine (0, 0, et 25%, respectivement). Une forte prévalence des gènes qnr augmente le taux de résistance croissant d'E. coli contre l'antibiotique quinolone à l'étude.

Keywords [French]

  • Qnr
  • antibiotiques quinolones
  • E. coli
  • résistance à médiation plasmidique

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Archives of Razi Institute
Volume 76, Issue 3
September 2021
Pages 561-573

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