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Author Spotlight: Advancing Antibiotic Resistance Research Using an Efflux-Deficient Bacterial Strain and a Single-Copy Gene Expression System
Published on: January 5, 2024
Detection of efflux pump genes in multiresistant Acinetobacter baumannii ST2 in Iran
Zahra Meshkat1, Himen Salimizand2,3, Yousef Amini4,5
11Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
Abstract:
Acinetobacter baumannii, as a nosocomial pathogen has become a worldwide concern in recent years. In the current study, the resistance to tetracyclines and colistin were assessed in the isolates from different provinces of Iran.During the timeline of this study, a number of 270 isolates of A. baumannii were collected from tracheal aspirates, wounds, urine and blood cultures. The minimum inhibitory concentration (MIC) for tetracycline, doxycycline, minocycline, tigecycline and colistin were evaluated. Tetracycline resistance genes were assessed by PCR. The mean expression level of adeB, adeJ and adeG were assessed using semi quantitative Real-Time PCR. The clonal relationship of the isolates was evaluated by the repetitive extragenic palindromic PCR (REP-PCR), International Clonal (IC) Lineage Multiplex PCR and multilocus sequence typing (MLST) (Pasteur scheme) methods.The MIC by microdilution method showed that 87.5, 51.4, 28, 0.74 and 0% of the isolates were resistant to tetracycline, doxycycline, minocycline, tigecycline and colistin respectively. The prevalence of tetracycline resistance genes was 99.2, 99.2, 98, 86.7, 10, 3.33, 0.37, 0% for adeB, adeJ, adeG, tetB, tetA(39), tetA, tetM and tetH in tetracycline-resistant isolates. Moreover, the expression level of adeB, adeJ, adeG genes in tigecycline-nonsusceptible A. baumannii (TNAB) strain was higher compared to the tigecycline-susceptible A. baumannii (TSAB). A broad genomic diversity was revealed, but ST2 was the most prevalent ST. Our results indicated that tetracycline resistance in Iran is mediated by resistance-nodulation-cell division (RND) and tetB efflux pumps.
Insights
High tetracycline resistance in Acinetobacter baumannii isolates in Iran is linked to efflux pumps, specifically resistance-nodulation-cell division (RND) and tetB. Colistin resistance was not observed in this study.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Acinetobacter baumannii is a significant global nosocomial pathogen.
- Understanding antimicrobial resistance patterns is crucial for effective treatment.
Purpose of the Study:
- To assess tetracycline and colistin resistance in Iranian A. baumannii isolates.
- To investigate the genetic mechanisms and clonal relationships underlying resistance.
Main Methods:
- Minimum Inhibitory Concentration (MIC) determination for various antibiotics.
- Polymerase Chain Reaction (PCR) for detecting tetracycline resistance genes.
- Semi-quantitative Real-Time PCR for efflux pump gene expression.
- Repetitive Extragenic Palindromic PCR (REP-PCR), International Clonal (IC) Lineage Multiplex PCR, and Multilocus Sequence Typing (MLST) for genetic relatedness.
Main Results:
- High resistance rates observed for tetracycline (87.5%), doxycycline (51.4%), and minocycline (28%).
- Tigecycline resistance was low (0.74%), and no colistin resistance was detected.
- Prevalence of resistance genes like adeB, adeJ, adeG, and tetB was high among resistant isolates.
- Increased expression of adeB, adeJ, and adeG genes in tigecycline-nonsusceptible strains.
- ST2 was the most prevalent sequence type, indicating broad genomic diversity.
Conclusions:
- Tetracycline resistance in Iran is primarily mediated by RND and tetB efflux pumps.
- Efflux pump gene expression is associated with reduced susceptibility to tigecycline.
- Genomic diversity exists, but specific clones like ST2 are prevalent.
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