Abstract
During the period from November 2009 to April 2010, 84 out of 560 extended spectrum β-lactamase (ESBL) producing negative bacteria were isolated from patients in different departments of the Ahmadi Hospital in Kuwait. The isolates were collected from urine catheter, wound, sputum, blood and other different samples. The ESBL infection rate in the in-patients was 62% and part of them (19%) were in the intensive care unit. All the isolated bacteria were identified and tested for antimicrobial susceptibility using an automated system (VITEK 2) and different antibiotic discs (15) by standard disc diffusion. The number of the recorded isolated multi-resistant Gram's negative bacteria was 54 isolates of Escherichia coli, 18 of Klebsiella pneumoniae, 11 of Pseudomonas aeruginosa, six of Proteus mirabilis, five of Enterobacter cloacae, four of Acinetobacter baumanii and one of Enterobacter aerogenes. They were resistant to the third generation of cephalosporins; Ceftazidime, Cefotaxime and Ceftriaxone. Meropenam (MEM) was the highest effective antibiotic against all the isolated bacteria (86%). The production of the ESBL was detected by phenotypic methods using E-test (96.4%), double disk synergy test (95%) and VITEK 2 (84.5%) in all multi-resistant isolates except A. baumanii and P. aeruginosa. All ESBL producing isolates were extracted and subjected to PCR using blaSHV, blaCTX-M and blaTEM primers. The bla-CTX-M (63.1%) was the most predominant ESBL gene that was produced in abundance by 42 isolates of E. coli. The most predominant ESBL isolates producing bla-TEM, bla-CTX-M and bla-SHV genes were successfully identified by 16 S rDNA. The conjugation assay between E. coli HB101 and the most predominant ESBL producing E. coli showed that the bla-CTX-M gene was able to be transferrable suggesting that they were plasmid mediated. Key words: Extended spectrum β-lactamase (ESBL), VITEK 2, E-Test, DDST, polymeric chain reaction (PCR), 16S rDNA.
Highlights
Extended spectrum of β -lactamases (ESBLs) are enzymes produced by a variety of Gram negative bacteria which confer an increased resistance to commonly used antibiotics
The number of the multi-resistant isolates was identified as E. coli (54 out of 358), K. pneumoniae (18 out of 96), P. aeruginosa (11 out 58), P. mirabilis (6 out of 17), E. cloacae (5 out of 18), A. baumanii (4 out of 9) and E. aerogenes
All the extended spectrum beta lactamases (ESBLs) producers were recovered from urine specimens and female patients were the predominant for ESBL producing E. coli infection
Summary
Extended spectrum of β -lactamases (ESBLs) are enzymes produced by a variety of Gram negative bacteria which confer an increased resistance to commonly used antibiotics. They are a worrying global public health issue as infections caused by such enzyme producing organisms are associated with a higher morbi-. The introduction of the third-generation of cephalosporins into clinical practice in the early 1980s is heralded as a major breakthrough in the fight against β -lactamase-mediated bacterial resistance to antibiotics These cephalosporins have been developed in response to the increased prevalence of ß-lactamases in certain organisms (for example, ampicillin hydrolyzing TEM-1 and SHV-1 β -lactamases in E. coli and K. pneumoniae). Thereafter, the number of ESBL variants occurring through amino acid mutations has progressively increased while demonstrating geographic variations (Winokur et al, 2001)
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