Molecular identification of blaSHV, blaLEN and blaOKP β-lactamase genes in Klebsiella pneumoniae by bi-directional sequencing of universal SP6- and T7-sequence-tagged blaSHV-PCR amplicons

Abstract

Molecular identification of bla SHV, bla LEN and bla OKP β-lactamase genes in Klebsiella pneumoniae by bi-directional sequencing of universal SP6-and T7-sequence-tagged bla SHV-PCR amplicons, 2009, Molecular and Cellular Probes, (23), 3-4, 195-200

Comparison of (GTG)5-oligonucleotide and ribosomal intergenic transcribed spacer (ITS)-PCR for molecular typing of Klebsiella isolates

“…Phenotypic ESBL-screening and susceptibility testing was performed on all (approximately 800 isolates/year) Klebsiella pneumoniae and Klebsiella oxytoca clinical isolates collected during 2001-and spring 2007 at the Department of Clinical Microbiology, University Hospital Linköping, Sweden with agar disk diffusion according to the Swedish Reference Group for Antibiotics (http://www.srga.org) as described previously (Monstein et al, 2007;Tärnberg et al, 2009). In brief, Cefadroxil was used for the screening of cephalosporin resistance, which was followed up by testing of resistant isolates with cefotaxime and ceftazidime or direct testing with cefotaxime and ceftazidime with disk diffusion and Etest (bioMerieux Sverige AB, Askim, Sweden).…”

“…Nineteen K. pneumoniae and 35 K. oxytoca isolates were selected and stored in glycerol containing Nutrient-broth No 2 (Lab M, Bury, UK) at -70° C until analysis Tärnberg et al, 2009 …”

“…To perform concurrent genotyping analysis omitting multiple bacterial culturing, sufficient amounts of bacterial DNA were produced by multiple displacement amplification as described elsewhere (Monstein et al, 2005;Monstein et al, 2009;Tärnberg M, et al, 2009). …”

“…3). Tärnberg et al, 2009). ESBL-genotypes in R. planticola (PLA) and R. ornithinolytica (ORN) type strains have been described earlier (Walckenaer et al, 2004).…”

“…I-II indicates two major cluster representing K. pneumoniae and K. oxytoca strains, and III represents outliers at a similarity level of 60 %, respectively. Bacterial phenotypes, ESBL genotypes (CTX-M, K1, SHV, LEN and OKP) of the clinical isolates were established in previous studies Tärnberg et al, 2009). ESBL-genotypes in R. planticola (PLA) and R. ornithinolytica (ORN) type strains have been described earlier (Walckenaer et al, 2004).…”