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Abstract
Urinary tract infection is primarily caused by Escherichia coli. Multidrug resistance and their rapid dissemination in this pathogenic microbe complicate therapeutic strategies and threaten public health. Conjugation systems responsible for interbacterial transmission of antibiotic resistance are plasmid-encoded and can be classified as the P, F, and I types. Specific pili types and pili associated proteins were related to the transfer among this gram-negative organism and were thought to depend on contacts created by these structures at the time of DNA transport. In this study, conjugation system types of the plasmids that harbor multidrug resistant genes (aac-1b-cr, oqxAB, qnrB, qnrS, blaTEM, blaOXA) amongst 19 E. coli uropathogenic isolates were characterized under ciprofloxacin/ceftazidime selection individually by pili and pili associated gene types. Investigations indicated incidence of single plasmid of multiple replicon type amongst the transconjugants. blaTEM, blaCTX–M, blaOXA, aac-1b-cr, oqxAB, qnrB, qnrS genes in varied combination were observed to be successfully co-transmitted against ceftazidme/ciprofloxacin selection. Seven primer pair sets were selected that encodes pili and pili associated genes (traF, trwJ, traE, trhE, traG, pilM, pilx4) by nucleotide database search tools using annotated plasmids of different incompatibility types to assign the conjugation system type of the transmissible resistant plasmids by PCR. traF was predominant irrespective of drug selection that indicated F-type conjugation system was responsible for transmission of resistant plasmids which results in the rapid dissemination of antibiotic resistance in the isolates screened. Therefore this is a first report of its kind that investigated pili and pili associated genes to bio-type multidrug resistant plasmids and their transmission in clinical settings amongst uropathogenic E. coli circulated in the eastern part of India.
Highlights
Urinary tract infection (UTI) is known to cause significant levels of morbidity and mortality in developed countries and has become a public health concern
In the present study 50 urine samples yielded significant growth out of 80 samples collected from patients suffering from UTI. 19 E. coli isolates were identified from the 50 urine culture positive samples by routine biochemical analysis
All 19 E. coli isolates were resistant toward ciprofloxacin, ceftazidime, cefotaxime, cefipime, amikacin, levofloxacin, 1http://blast.ncbi.nlm.nih.gov/ 2https://www.ncbi.nlm.nih.gov/nucleotide/ 3http://www.ebi.ac.uk/Tools/msa/clustalw2/ 4http://primerdigital.com/fastpcr.html nitrofurantoin, and imepenem except 2, 3, 4, 8, and 8 out of the 19 which showed intermediate resistance against amikacin, levofloxacin, nitrofurantoin and imepenem, respectively
Summary
Urinary tract infection (UTI) is known to cause significant levels of morbidity and mortality in developed countries and has become a public health concern. Emergence of antibiotic resistance in this bacterial pathogen is recognized as one of the greatest threats to global healthcare management system (reviewed in Hadifar et al, 2017). Multidrug resistant (MDR) isolates causing UTI have serious implications for the empiric therapy against pathogenic isolates and for the possible co-selection of antimicrobial resistant pathogens. There are several factors responsible for dissemination of antimicrobial resistance genes among these pathogens, and plasmid-mediated transfer has been considered one of the most important mechanisms for the horizontal transfer of multidrug resistance (Akingbade et al, 2014). The most widely used plasmid classification scheme is PCR based replicon typing (PBRT) which exploited loci encoding plasmid replication machinery (Carattoli et al, 2005, 2014) which provided insights into resistance plasmid epidemiology, such as whether resistance dissemination involves diverse plasmids or one dominant “epidemic” type (Valverde et al, 2009)
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