Molecular characteristics of extended-spectrum β-lactamases in clinical isolates of Enterobacteriaceae from the Philippines.
β-Lactamases, including extended-spectrum β-lactamases (ESBLs) and AmpC β-lactamases, are major resistance mechanisms of Enterobacteriaceae. Emergence of plasmid-mediated quinolone resistance (PMQR) determinants in ESBL-producing isolates poses a global threat. The molecular characterisitcs of ESBL and PMQR determinants in the Philippines are not well characterized. In this study, we investigated ESBLs and AmpC β-lactamases in clinical isolates of Enterobacteriaceae from the Philippines, and analyzed the association between ESBL and PMQR genes. A total of 91 amoxicilin non-susceptible Enterobacteriaceae were collected at the Research Institute for Tropical Medicine of the Philippines from 2006 to 2008. AmpC- or ESBL-producing isolates were screened by detecting a zone diameter for cefoxitin ≤ 14 mm or cefpodoxime ≤ 20 mm, respectively. Possible ESBL-producing strains were assessed by the ESBL confirmation test of the Clinical and Laboratory Standards Institute. PCR and sequencing were performed to detect the ESBL and PMQR genes. The number of ESBL-producers and AmpC-producers confirmed phenotypically was 17 (18.7%) and 61 (67.0%), respectively. Of 17 phenotypic ESBL-producers, 14 isolates had ESBL genes, including 6 of Escherichia coli, 3 of Enterobacter cloacae, 2 of Enterobacter aerogenes, 2 of Klebsiella pneumoniae, and 1 of Klebsiella ozaenae. Among these isolates, there were 13, 4, and 12 with bla(CTX-M), bla(SHV), and bla(TEM), respectively. Of the bla(CTX-M)-positive isolates, bla(CTX-M-15) shows the highest prevalence, followed by bla(CTX-M-3) and bla(CTX-M-14). Of 14 ESBL-producers identified by PCR, 4, 6, and 7 isolates were positive for qnrB, qnrS, and aac(6')-Ib-cr, respectively. The frequency of aac(6')-Ib-cr positivity was significantly higher among CTX-M-15-producing isolates. Thus, we identified bla(CTX-M), aac(6')-Ib-cr, and qnr in ESBL-producing Enterobacteriaceae from the Philippines, and revealed a significant association between bla(CTX-M-15) and aac(6')-Ib-cr. Local epidemiological data are important for implementing appropriate antimicrobial therapy and effective infection control measures. Continuous monitoring of antimicrobial resistance genes in the Philippines will be required.
Kanamori H
，Navarro RB
，Yano H
，Sombrero LT
，Capeding MR
，Lupisan SP
，Olveda RM
，Arai K
，Kunishima H
，Hirakata Y
，Kaku M
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Molecular characteristics of extended-spectrum beta-lactamase-producing Escherichia coli from the Chicago area: high prevalence of ST131 producing CTX-M-15 in community hospitals.
This study was designed to characterise 30 non-duplicate extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli clinical isolates from the community in the Chicago metropolitan area collected during 2008. The majority of isolates (n=28) were recovered from urine and 2 isolates were from blood. Molecular characterisation was done using the following techniques: isoelectric focusing; polymerase chain reaction (PCR) and sequencing of bla(ESBL); PCR for plasmid-mediated quinolone resistance determinants; identification of ST131; phylogenetic grouping; and replicon typing. Genetic relatedness was determined by pulsed-field gel electrophoresis (PFGE) with XbaI and repetitive sequence-based PCR (rep-PCR) typing. Twenty-six (87%) of the ESBL-producing E. coli were positive for bla(CTX-M) genes (22 CTX-M-15 and 4 CTX-M-14), whilst the remaining 4 isolates produced SHV-2. Twenty-eight isolates (93%) were non-susceptible to ciprofloxacin and 16 (53%) were positive for aac(6')-Ib-cr. Overall, 16 (53%) of the ESBL-producers belonged to clonal complex ST131 that produced CTX-M-15 or CTX-M-14. Molecular characteristics of ST131 showed that it belonged to three distinct but related PFGE clones, was derived from phylogenetic group B2 and contained IncFII type plasmids. These results illustrate that E. coli clonal complex ST131 producing CTX-M-15, CTX-M-14, OXA-1, TEM-1 and aac(6')-Ib-cr has emerged as an important cause of community-onset urinary tract infections caused by ESBL-producing E. coli isolates in the Chicago area.
Peirano G
，Costello M
，Pitout JD
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