Open AccessSimple and reliable multiplex PCR assay for detection of bla TEM , bla SHV and bla OXA‐1 genes in Enterobacteriaceae
Author(s) -
Colom Karmele,
Pérez Javier,
Alonso Rodrigo,
FernándezAranguiz Agueda,
Lariño Eva,
Cisterna Ramón
Publication year - 2003
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/s0378-1097(03)00306-9
Subject(s) - enterobacteriaceae , multiplex polymerase chain reaction , biology , microbiology and biotechnology , cefotaxime , escherichia coli , polymerase chain reaction , plasmid , clavulanic acid , multiplex , cephalosporin , gene , amoxicillin , genetics , antibiotics
Third‐generation cephalosporin resistance is often mediated by TEM‐ and SHV‐type β‐lactamases in Enterobacteriaceae. TEM‐type and OXA‐1 enzymes are the major plasmid‐borne β‐lactamases implicated in amoxicillin–clavulanic acid resistance in Escherichia coli isolates. We have developed a rapid and simple multiplex polymerase chain reaction (PCR) which discriminates bla TEM , bla SHV and bla OXA‐1 genes by generating fragments of 516, 392 and 619 bp respectively. Multiplex PCR analysis of 51 amoxicillin–clavulanate resistant E. coli isolates detected bla TEM and bla SHV genes in 45 and two strains, respectively, and only one strain harboured a bla OXA‐1 gene. Twenty‐three of the 40 cefotaxime‐resistant Enterobacteriaceae isolates produced amplicons with a size compatible with the presence of bla TEM (13 strains), bla SHV (six strains) genes or the association of both genes (four strains). These results were verified by colony hybridisation. Therefore, multiplex PCR is a suitable tool for initial rapid screening of bla genes in Enterobacteriaceae.