Molecular characterisation by PCR-restriction fragment length polymorphism of TEM beta-lactamases
- PMID: 8586268
- DOI: 10.1111/j.1574-6968.1995.tb07938.x
Molecular characterisation by PCR-restriction fragment length polymorphism of TEM beta-lactamases
Abstract
To rapidly characterise TEM-derived extended-spectrum beta-lactamases a fast and easy method using polymerase chain reaction-restriction fragment length polymorphism was developed. This method was validated with ten reference TEM-type extended-spectrum beta-lactamases. The mutations involved in TEM-20 and TEM-21, which were previously reported only with biochemical analysis, were then characterised. TEM-20 differed from TEM-19 by a silent mutation at position 925 (A for G), and TEM-21 differed from TEM-3 and TEM-14 by a single mutation (G for A) in an unreported position 660. beta-lactamase conferring low resistance to ceftazidime (TEM-29), was described. TEM-29 derived from TEM-1, with an amino acid substitution, his-164. Finally, the combination of polymerase chain reaction-restriction fragment length polymorphism and plasmid analysis allowed us to investigate nosocomial outbreaks due to clinical isolates of multi-resistant Klebsiella pneumoniae in three hospitals.
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