Molecular characterisation by PCR-restriction fragment length polymorphism of TEM beta-lactamases

FEMS Microbiol Lett. 1995 Dec 15;134(2-3):203-8. doi: 10.1111/j.1574-6968.1995.tb07938.x.


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.

MeSH terms

  • Cross Infection / epidemiology
  • Cross Infection / microbiology
  • DNA, Bacterial / genetics
  • Disease Outbreaks
  • Drug Resistance, Multiple / genetics
  • Humans
  • Klebsiella Infections / epidemiology
  • Klebsiella Infections / microbiology
  • Klebsiella pneumoniae / drug effects
  • Klebsiella pneumoniae / enzymology*
  • Klebsiella pneumoniae / genetics*
  • Plasmids / genetics
  • Point Mutation
  • Polymerase Chain Reaction
  • Polymorphism, Restriction Fragment Length
  • beta-Lactamases / chemistry
  • beta-Lactamases / genetics*


  • DNA, Bacterial
  • beta-Lactamases