Preprint Article Version 1 This version is not peer-reviewed

Extended Spectrum Beta-Lactamase (ESBL)-Mediated Resistance to Third Generation Cephalosporins and Conjugative Transfer of Resistance in Gram-Negative Bacteria Isolated from Hospitals in Tamil Nadu, India

Version 1 : Received: 8 October 2019 / Approved: 9 October 2019 / Online: 9 October 2019 (11:03:44 CEST)

How to cite: Ramesh , N.; Rajesh Kannan, V.; Bozdogan, B.; Krishnakumar, V.; Manohar, P. Extended Spectrum Beta-Lactamase (ESBL)-Mediated Resistance to Third Generation Cephalosporins and Conjugative Transfer of Resistance in Gram-Negative Bacteria Isolated from Hospitals in Tamil Nadu, India. Preprints 2019, 2019100103 (doi: 10.20944/preprints201910.0103.v1). Ramesh , N.; Rajesh Kannan, V.; Bozdogan, B.; Krishnakumar, V.; Manohar, P. Extended Spectrum Beta-Lactamase (ESBL)-Mediated Resistance to Third Generation Cephalosporins and Conjugative Transfer of Resistance in Gram-Negative Bacteria Isolated from Hospitals in Tamil Nadu, India. Preprints 2019, 2019100103 (doi: 10.20944/preprints201910.0103.v1).

Abstract

Clinical pathogens especially Gram-negative bacteria developing resistance to third-generation cephalosporins are making the clinical outcome more complicated and serious. This study was undertaken to evaluate the distribution of extended-spectrum beta-lactamases in Tamil Nadu regions in India. For this study, clinical samples were collected from five different hospitals located in Tamil Nadu and ESBL producing Gram-negative isolates were characterized. Minimal inhibitory concentration (MIC) was performed using cefotaxime and ceftazidime. The blaESBL producing genes were screened using multiplex PCR for the genes, CTX-M group-1,-2,-8,-9,-26. Conjugation studies were performed using E. coli AB1157 as a recipient for the isolates harbouring plasmid-borne resistance following broth-mating experiment. In total, 1500 samples were collected and 599 Gram-negative bacteria were isolated that included Escherichia coli (n=233), Klebsiella pneumoniae (n=182), Pseudomonas aeruginosa (n=79), Citrobacter spp. (n=30), Proteus mirabilis (n=28), Salmonella spp. (n=21), Acinetobacter baumannii (n=12), Serratia spp. (n=6), Shigella spp. (n=4), Morganella morganii (n=3) and Providencia spp. (n=1). MIC results showed that 358 isolates were resistant to cefotaxime and ceftazidime. Further, ESBL gene amplification results showed that 19 isolates had CTX-M group-1 gene including E. coli (n=16), K. pneumoniae (n=2) and P. aeruginosa (n=1) whereas one M. morganii isolate had CTX-M group-9 gene in their plasmid. Through conjugation studies, 12/20 isolates were found to be involved in the transformation of its plasmid-borne resistance gene. Our study highlighted the role of horizontal gene transfer in the dissemination of plasmid-borne blaCTX-M resistance genes among ESBL producing isolates.

Subject Areas

beta-lactamase; cephalosporin; cefotaxime; transconjugation; plasmid-borne resistance

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