Molecular characterisation by PCR-restriction fragment length polymorphism of TEM beta-lactamases.
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.
Arlet
G
G
Service de Microbiologie, Hôpital Saint-Louis, Paris, France.
Brami
G
G
Décrè
D
D
Flippo
A
A
Gaillot
O
O
Lagrange
P H
PH
Philippon
A
A
D003428:Cross Infection / Q000453:epidemiology / Q000382:microbiology; D004269:DNA, Bacterial / Q000235:genetics; D004196:Disease Outbreaks; D018432:Drug Resistance, Multiple / Q000235:genetics; D006801:Humans; D007710:Klebsiella Infections / Q000453:epidemiology / Q000382:microbiology; D007711:Klebsiella pneumoniae / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D001618:beta-Lactamases / Q000737:chemistry / Q000235:genetics*
D004269:DNA, Bacterial; D001618:beta-Lactamases