Molecular characteristics of extended-spectrum beta-lactamases among gram-negative isolates collected in Cairo University Hospital

Phenotyping is commonly used for detection of extended-spectrum beta-lactamase (ESBL) production in gram-negative isolates. ESBLs are mainly coded for by four important genes, namely bla (TEM), bla (SHV), bla (CTX-M), and bla (OXA). Our aim in this study is to assess use of a multiplex PCR as a rapid method to identify four common genes responsible for ESBL production in different gram-negative isolates. All 793 clinical isolates are subjected to both screen and confirmatory testing for ESBL production using double disc synergy testing (DDST). Two hundred isolates with the ESBL phenotype are subjected to multiplex PCR for detection of the four genes bla (TEM, SHV, CTX-M, and OXA). The isolates were obtained from various clinical specimens: 68 (34 %) were isolated from urine cultures, 43 (21.5 %) from sputum, 26 (13 %) from wounds, 34 (17 %) from blood culture, 20 (10 %) from stool of healthy carrier and nine (4.5 %) from bronchoalveolar lavages. In this study, 83 isolates (41.5 %) were from outpatients (urine and stool specimens only), and the remaining 117 isolates (58.5) were from inpatients. By PCR technique, 181 isolates were found to be ESBL producers. blaTEM was the commonest genotype (39.2 %), followed by blaSHV (32.5 %) and blaCTX-M (30.9 %), either alone or in combination. Acinetobacter baumannii isolate had none of the ESBL genes. Eighteen (9.9 %) out of 181 isolates carried more than one type of beta-lactamase genes. Our study demonstrated rapid detection of bla (TEM, SHV, CTX-M, and OXA) in isolates belonging to Enterobacteriaceae and other nonfermenting clinical isolates using multiplex PCR. This genotypic method provided a rapid and efficient differentiation of ESBLs in the laboratory.