Abstract

IntroductionThe occurrence of multiple β-lactamases among bacteria only limits the therapeutic options but also poses a challenge. A study using boronic acid (BA), an AmpC enzyme inhibitor, was designed to detect the combined expression of AmpC β-lactamases and extended-spectrum β-lactamases (ESBLs) in bacterial isolates further different phenotypic methods are compared to detect ESBL and AmpC.MethodsA total of 259 clinical isolates of Enterobacteriaceae were isolated and screened for ESBL production by (i) CLSI double-disk diffusion method (ii) cefepime- clavulanic acid method (iii) boronic disk potentiation method. AmpC production was detected using cefoxitin alone and in combination with boronic acid and confirmation was done by three dimensional disk methods. Isolates were also subjected to detailed antibiotic susceptibility test.ResultsAmong 259 isolates, 20.46% were coproducers of ESBL and AmpC, 26.45% were ESBL and 5.40% were AmpC. All of the 53 AmpC and ESBL coproducers were accurately detected by boronic acid disk potentiation method.ConclusionThe BA disk test using Clinical and Laboratory Standards Institute methodology is simple and very efficient method that accurately detects the isolates that harbor both AmpCs and ESBLs.

Highlights

  • The occurrence of multiple β-lactamases among bacteria only limits the therapeutic options and poses a challenge

  • Among 259 clinical isolates of Enterobacteriaceae, 68 (26.25%) and 14(5.4%) were pure ESBL and AmpC producers respectively; 53 (20.46%) isolates were combined ESBL and AmpC producers; and 124 (47.87%) of the isolates did not harbor any type of enzyme (Table 1)

  • The rate of detection of ESBLs by the Clinical Laboratory Standards Institute (CLSI) confirmatory test for clinical isolates that produce both ESBLs and AmpC (20.46%) was lower than that for clinical isolates that produce ESBLs but not AmpC (26.45%)

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Summary

Introduction

The occurrence of multiple β-lactamases among bacteria only limits the therapeutic options and poses a challenge. Conclusion: The BA disk test using Clinical and Laboratory Standards Institute methodology is simple and very efficient method that accurately detects the isolates that harbor both AmpCs and ESBLs. The rapid global dissemination of Enterobacteriaceae harboring plasmid-borne extended-spectrum β-lactamases (ESBLs) and plasmid-mediated AmpC β-lactamases represents a significant clinical threat [1,2]. ESBLs producing organism confer resistance to penicillin, cephalosporins, and monobactams They cannot hydrolyze cephamycins and are inhibited by clavulanic acid (CA) [3]. Inappropriate use of cephalosporins in clinical practice led to the emergence of bacteria producing multiple βlactamases This leads to therapeutic failure when β-lactam drugs or β-lactam/inhibitor combination are used [6 ]. The present study was aimed to evaluate the usage of boronic acid in a phenotypic confirmatory test to detect ESBL among AmpC βlactamases producing isolates

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