Detection of Plasmid-mediated Ampc β-lactamases Among E.coli and Klebsiella pneumoniae by Multiplex PCR”

  • Anuradha Basavaraju
  • Praveena Muttaraju Mamata Medical College , affiliated to NTR university of health sciences
Keywords: AmpC beta-lactamase, ESBL, E.coli, Klebsiella, PCR

Abstract

Background: Gram negative bacteria are acquiring drug resistance due to Extended spectrum beta lactamase (ESBL) production and also Plasmid mediated AmpC beta-lactamases (PMABLs). This is one of the major causes of multi-drug resistance among E.coli and Klebsiellainclinical practice. Detection of PMABL genes by molecular methods such as multiplex PCR gives accurate results in specific identification.Methods: ESBL producing strains of 40 E.coli and Klebsiella were tested phenotypically for Plasmid mediated AmpC beta-lactamase production by using cefoxitin disk. The genes coding for PMABLs production was tested by multiplex PCR. Antibiotic susceptibility pattern of the isolates was also tested.Results:  22(55%) of E.coli and 17(42.5%) of Klebsiella pneumoniae were phenotypically producing AmpC beta-lactamases. On genotypic testing 15(37.5%) E.coli and 11(28%) Klebsiella pneumoniae were positive for plasmid mediated AmpC beta-lactamases. Plasmid encoded AmpC genes in E.coli are CIT/EBC, CIT, and EBC. In Klebsiella pneumoniae the genes were CIT/DHA, CIT, and DHA. All the isolates showed 100% resistance to Cefoxitin and amox/clav and also higher degrees of resistance to cefotaxime, ceftazidime, cefepime, aztreonam and piperacillin/ tazobactam.Conclusion: ESBL producing strains of E.coli and Klebsiella are developing drug resistance due to the production of PMABLs. Detection of genes coding for PMABL production are best tested by multiplex PCR which gives accurate results than phenotypic detection methods.

Author Biography

Praveena Muttaraju, Mamata Medical College , affiliated to NTR university of health sciences
Department of Microbiology, Mamata Medical College

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Published
2016-08-24
Section
Original Article