Restriction fragment length dimorphism-PCR method for the detection of extended-spectrum β-lactamases unrelated to TEM- and SHV-types

The diagnostic ability of the restriction fragment length dimorphism-polymerase chain reaction (RFLD-PCR) method was evaluated. Seven primer pairs, newly designed from 44 beta -lactamase genes encoding extended-spectrum beta -lactamases not related to TEM- and SHV-types, were used to differentiate OXA-2, FOX-3, CMY-3, IMP-1, and IMI-1 beta -lactamases. The RFLD-PCR a as carried out successfully. and these genes were differentiated by the sizes of their PCR products and by the difference in restriction fragment length when each amplicon was digested with a unique restriction enzyme. This discriminatory detection of the genes was confirmed by sequencing the PCR products. (C) 2001 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.