The molecular characteristics of cefepime-susceptible Escherichia coli and Klebsiella spp. isolates with a positive β-lactamase screening test result but negative confirmation

Abstract

A negative extended-spectrum β-lactamase (ESBL) phenotypic confirmation test result obtained after a positive ESBL screening test result but which was cefepime-susceptible (NCPSCS) using the Clinical and Laboratory Standards Institute (CLSI) methods has been observed among isolates of Escherichia coli and Klebsiella spp. in the antimicrobial surveillance program in Shanghai, China. Among isolates collected from Huashan Hospital in 2005, NCPSCS strains were observed in 2.5% of 433 E. coli isolates and in 1.2% of 562 Klebsiella spp. isolates. We then selected 11 E. coli isolates and seven Klebsiella spp. NCPSCS isolates. Transmission electron microscopy (TEM), SHV, plasmid-borne AmpC, and CTX-M type ESBL genes were detected by the polymerase chain reaction (PCR) method. We found that all except two K. pneumoniae strains of NCPSCS isolates producing ESBL and AmpC harbored a plasmid-borne CMY-2 or DHA-1 type AmpC enzyme. The majority of NCPSCS E. coli and K. pneumoniae strains from Shanghai harbor plasmid-borne AmpC enzyme, and we recommend that, when NCPSCS strains are identified, further work such as the PCR detection of ESBL genes is necessary to determine whether they will produce ESBLs. The ESBL-positive strains should be reported as resistant to cefepime according to the CLSI guidelines.