Multiplex PCR amplification assay for the detection of blaSHV, blaTEM and blaCTX-M genes in Enterobacteriaceae.
Extended-spectrum beta-lactamases (ESBLs) are often mediated by (bla-)SHV, (bla)TEM and (bla)CTX-M genes in Enterobacteriaceae and other Gram-negative bacteria. Numerous molecular typing methods, including PCR-based assays, have been developed for their identification. To reduce the number of PCR amplifications needed we have developed a multiplex PCR assay which detects and discriminates between (bla-)SHV, (bla)TEM and (bla)CTX-M PCR amplicons of 747, 445 and 593 bp, respectively. This multiplex PCR assay allowed the identification of (bla-)SHV, (bla)TEM and (bla)CTX-M genes in a series of clinical isolates of Enterobacteriaceae with previously characterised ESBL phenotype. The presence of (bla)SHV, (bla)TEM and (bla)CTX-M genes was confirmed by partial DNA sequence analysis. Apparently, the universal well-established CTX-M primer pair used here to reveal plasmid-encoded (bla)CTX-M genes would also amplify the chromosomally located K-1 enzyme gene in all Klebsiella oxytoca strains included in the study.
Monstein
H-J
HJ
Division of Clinical Microbiology, University Hospital Linköping, Linköping, Sweden.
Ostholm-Balkhed
A
A
Nilsson
M V
MV
Nilsson
M
M
Dornbusch
K
K
Nilsson
L E
LE
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis*; D004755:Enterobacteriaceae / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D010641:Phenotype; D016133:Polymerase Chain Reaction / Q000379:methods*; D001618:beta-Lactamases / Q000032:analysis* / Q000235:genetics*
D017931:DNA Primers; D004269:DNA, Bacterial; D001618:beta-Lactamases