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Journal of Clinical Microbiology, April 2003, p. 1463-1468, Vol. 41, No. 4
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.4.1463-1468.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Characterization of Clinical Isolates of Enterobacteriaceae from Italy by the BD Phoenix Extended-Spectrum ß-Lactamase Detection Method

Maurizio Sanguinetti,1* Brunella Posteraro,1 Teresa Spanu,1 Daniela Ciccaglione,1 Lucio Romano,1 Barbara Fiori,1 Giuseppe Nicoletti,2 Stefania Zanetti,3 and Giovanni Fadda1

Istituto di Microbiologia, Università Cattolica del Sacro Cuore, Rome,1 Dipartimento di Scienze Microbiologiche e Ginecologiche, Sezione di Microbiologia, Università di Catania, Catania,2 Dipartimento di Scienze Biomediche, Sezione di Microbiologia Sperimentale e Clinica, Università di Sassari, Sassari, Italy3

Received 13 August 2002/ Returned for modification 5 December 2002/ Accepted 30 December 2002

Production of extended-spectrum ß-lactamases (ESBLs) is an important mechanism of ß-lactam resistance in Enterobacteriaceae. Identification of ESBLs based on phenotypic tests is the strategy most commonly used in clinical microbiology laboratories. The Phoenix ESBL test (BD Diagnostic Systems, Sparks, Md.) is a recently developed automated system for detection of ESBL-producing gram-negative bacteria. An algorithm based on phenotypic responses to a panel of cephalosporins (ceftazidime plus clavulanic acid, ceftazidime, cefotaxime plus clavulanic acid, cefpodoxime, and ceftriaxone plus clavulanic acid) was used to test 510 clinical isolates of Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Proteus mirabilis, Providencia stuartii, Morganella morganii, Enterobacter aerogenes, Enterobacter cloacae, Serratia marcescens, Citrobacter freundii, and Citrobacter koseri. Of these isolates, 319 were identified as ESBL producers, and the remaining 191 were identified as non-ESBL producers based on the results of current phenotypic tests. Combined use of isoelectric focusing, PCR, and/or DNA sequencing demonstrated that 288 isolates possessed blaTEM-1- and/or blaSHV-1-derived genes, and 28 had a blaCTX-M gene. Among the 191 non-ESBL-producing isolates, 77 isolates produced an AmpC-type enzyme, 110 isolates possessed TEM-1, TEM-2, or SHV-1 ß-lactamases, and the remaining four isolates (all K. oxytoca strains) hyperproduced K1 chromosomal ß-lactamase. The Phoenix ESBL test system gave positive results for all the 319 ESBL-producing isolates and also for two of the four K1-hyperproducing isolates of K. oxytoca. Compared with the phenotypic tests and molecular analyses, the Phoenix system displayed 100% sensitivity and 98.9% specificity. These findings suggest that the Phoenix ESBL test can be a rapid and reliable method for laboratory detection of ESBL resistance in gram-negative bacteria.

* Corresponding author. Mailing address: Istituto di Microbiologia, Università Cattolica del S. Cuore, L.go F. Vito, 1, 00168 Rome, Italy. Phone: 39-6-30154964. Fax: 39-6-3051152. E-mail: msanguinetti{at}rm.unicatt.it.

Journal of Clinical Microbiology, April 2003, p. 1463-1468, Vol. 41, No. 4
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.4.1463-1468.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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