Dissemination of CTX-M-15 β-lactamase genes carried on inc FI and FII plasmids among clinical isolates of escherichia coli in a university hospital in Istanbul, Turkey

The CTX-M-1 group was found in 86.8% of the Escherichia coli isolates from Istanbul. A subset study revealed all isolates carrying blaCTX-M-15 genes flanked by the insertion element ISEcp1. Plasmid typing of transconjugates carrying blaCTX-M-15 showed that most isolates belonged to the Inc/rep FII group but that one isolate also belonged to the FI group

Time kill-assays of antibiotic combinations for multidrug resistant clinical isolates of OXA-48 carbapenemase producing Klebsiella pneumoniae

Treatment of infections caused by OXA-48 carbapenemase producing multidrug-resistant isolates often necessitates combination therapy. In vitro effect of different antibiotic combinations against multidrug-resistant (MDR) Klebsiella pneumoniae isolates were evaluated in this study.Meropenem-tobramycin (MER+TOB), meropenem-ciprofloxacin (MER+CIP), colistin-mer-openem (COL+MER), colistin-ciprofloxacin (COL+CIP) and colistin-tobramycin (COL+TOB) combinations were tested by time kill-assays. Each antibiotic alone and in combination at their Cmax values were tested against 4 clinical K. pneumoniae isolates at 1, 2, 4, 6, 8, 12 and 24 h. Effect of colistin and its associations were also assessed at 30 min. Bactericidal activity was defined as >= 3log10 CFU mL-1 decrease compared with initial inoculum. Synergy was defined as >= 2log10CFU mL-1 decrease by the combination compared with the most active single agent. Presence of blaOXA-48, blaNDM, blaVIM, blaIMP, blaKPC and blaCTX-M-1 genes was screened by PCR using specific primers.The blaOXA-48 gene was identified together with blaCTXM-1 group gene in all isolates. COL+MER demonstrated to be synergistic and bactericidal. MER+TOB showed synergistic and bactericidal effect on two strains although, regrowth was seen on other two strains at 24 h. MER+CIP exhibited indif-ferent effect on the strains.Combination therapy could be a potential alternative to treat MDR K. pneumoniae infections. This combination might prevent resistance development and secondary effects of colistin monotherapy. MER+TOB and MER+CIP might have an isolate-dependent effect, that may not always result in synergism

Investigation of metallo-beta-lactamase producing strains of Pseudomonas aeruginosa and Acinetobacter baumannii by E-test, disk synergy and PCR

Carbapenem non-susceptible Pseudomonas aeruginosa and Acinetobacter baumannii strains were tested for the presence of metallo-beta-lactamases (MBLs) by EDTA-synergy screening. Imipenem hydrolysis was investigated by a bioassay and IMP-/VIM-encoding genes by PCR. No blaIMP/VIM related genes or imipenemase activity were detected although E-test found all strains as MBL-positive. Disk synergy tests with 0.5M EDTA determined 63.6-100%, while those with 0.1M EDTA detected 0-7.7% of isolates as MBL producers. Most strains were susceptible to EDTA. In conclusion, for MBL-screening purposes, EDTA-synergy results change with molarity of EDTA, but even if some false positives are encountered, 0.1M EDTA seems to be acceptable

Characterization of Resistance Genes and Polymerase Chain Reaction-Based Replicon Typing in Carbapenem-Resistant Klebsiella pneumoniae

Background: Fifty isolates of Klebsiella pneumoniae isolated from clinical samples between 2012 and 2016 that were found to be resistant to carbapenems were included in this study. Materials and Methods: Resistance genes were investigated by performing PCR. Plasmid typing was performed using PCR-based replicon typing. The clonal relationships between the strains were investigated using pulsed-field gel electrophoresis (PFGE). Results: OXA-48-type carbapenemase genes were detected in 86% (n = 43/50) of K. pneumoniae isolates, whereas NDM-type carbapenemase genes were detected in 14% (n = 7/50) of the isolates. bla(TEM) was detected 60% (n = 30) of the strains, bla(SHV) in 78% (n = 39), bla(CTX-M-1) in 48% (n = 24), and bla(CTX-M-2)-type beta-lactamase in 10% (n = 5). bla(CTX-M-1) and bla(SHV) were concomitantly distributed in 40% (n = 20) of the strains, bla(TEM) and bla(SHV) in 54% (n = 27), bla(TEM), bla(SHV), and bla(CTX-M-1) in 32% (n = 16) and bla(CTX-M-1) and bla(CTX-M-2) in 10% (n = 5). Strain numbers 66, 69, 76, 77, and 78 coproduced carbapenemases, bla(CTX-M-1) and bla(CTX-M-2) in addition to bla(OXA-48) or bla(NDM-1) that were described as hybrid strains. IncR-type replicon was found in 50% (n = 25) of 50 isolates with plasmid typing, whereas IncA/C-type replicon was detected in 40% (n = 20) and IncFIIK-type replicon in 18% (n = 9) of the isolates. Outcomes of the transformation experiments showed that the OXA-48 gene was carried to the receiver cell on FII plasmids. No dominant epidemic clone was detected through PFGE. Conclusion: OXA-48 carbapenemase was found to be the most prevalent type of enzyme in our hospital, and the presence of NDM-1-type carbapenemase-carrying strain and an increase in their rate were detected

Analysis of Virulence Factors and Antimicrobial Resistance in Salmonella Using Molecular Techniques and Identification of Clonal Relationships Among the Strains

A total of 50 Salmonella enterica strains were isolated from clinical samples from 2009 to 2012 and analyzed for the presence of virulence genes found in SPI-1, SPI-2, and plasmids. The distribution and frequency of the antimicrobial resistance genes and plasmids were revealed, and pulsed-field gel electrophoresis (PFGE) patterns were investigated. Five genes were identified from the seven strains with resistance or intermediate resistance to ampicillin: blaSHV-1 (present in six strains), qnrS1 (present in five strains), blaTEM-1 (present in three strains), blaCTX-M-1 (present in one strain), and qnrB1 (present in one strain). One trimethoprim-sulfamethoxazole-resistant strain was positive for sulI but negative for sulII. In addition, we detected TEM-1 and qnrS1 in one strain; SHV-1 and qnrS1 in two strains; TEM-1, SHV-1, CTX-M-1, and qnrS1 in one strain; TEM-1, SHV-1, and qnrB1 in one strain; and SHV-1 and sulI genes in one strain together. Plasmid-based replicon typing assay revealed that all 50 strains carried FIIS, 13 carried I1, 1 carried I2, 4 carried P, 1 carried A/C, and 4 carried X1 replicon. PFGE was used to type 46 of the 50 strains and classify them into 22 major groups, 33 pulsotypes, and 8 major clusters. All strains carried all the virulence genes of interest on both Salmonella Pathogenicity Islands 1 and 2 and plasmids suggested high potential for pathogenicity. All antimicrobial-resistant strains contained at least one of the resistance genes of interest, confirming a phenotype-genotype association in antimicrobial resistance

Comparison of Four Phenotypic Assays and Check-Direct CPE for Detection of Carbapenemases-Producing Enterobacterales

Background: Early and accurate detection of carbapenemase-producing Enterobacterales (CPE) is fundamental to prevent their spread in hospital environment. Our objective was to compare between four commonly used phenotypic assays and Check-Direct CPE (CDCPE) multiplex PCR in CPE detection. We examined stool samples or rectal swabs for CPE, samples collected from 23 Jan 2017 to 23 Jul 2017 from patients in intensive-care units (ICUs) of our hospital

Comparison of the Novel Oxa-48 and Kpc K-SeT Assay, and Blue-Carba Test for the Detection of Carbapenemase-Producing Enterobacteriaceae Using PCR as a Reference Method

Fifty isolates of Klebsiella pneumoniae isolated from clinical samples between 2012 and 2016 that were found to be resistant to carbapenems were included in this study. Materials and Methods: Resistance genes were investigated by performing PCR. Plasmid typing was performed using PCR-based replicon typing. The clonal relationships between the strains were investigated using pulsed-field gel electrophoresis (PFGE). Results: OXA-48-type carbapenemase genes were detected in 86% (n = 43/50) of K. pneumoniae isolates, whereas NDM-type carbapenemase genes were detected in 14% (n = 7/50) of the isolates. bla(TEM) was detected 60% (n = 30) of the strains, bla(SHV) in 78% (n = 39), bla(CTX-M-1) in 48% (n = 24), and bla(CTX-M-2)-type beta-lactamase in 10% (n = 5). bla(CTX-M-1) and bla(SHV) were concomitantly distributed in 40% (n = 20) of the strains, bla(TEM) and bla(SHV) in 54% (n = 27), bla(TEM), bla(SHV), and bla(CTX-M-1) in 32% (n = 16) and bla(CTX-M-1) and bla(CTX-M-2) in 10% (n = 5). Strain numbers 66, 69, 76, 77, and 78 coproduced carbapenemases, bla(CTX-M-1) and bla(CTX-M-2) in addition to bla(OXA-48) or bla(NDM-1) that were described as hybrid strains. IncR-type replicon was found in 50% (n = 25) of 50 isolates with plasmid typing, whereas IncA/C-type replicon was detected in 40% (n = 20) and IncFIIK-type replicon in 18% (n = 9) of the isolates. Outcomes of the transformation experiments showed that the OXA-48 gene was carried to the receiver cell on FII plasmids. No dominant epidemic clone was detected through PFGE. Conclusion: OXA-48 carbapenemase was found to be the most prevalent type of enzyme in our hospital, and the presence of NDM-1-type carbapenemase-carrying strain and an increase in their rate were detected

Resistance to macrolide, lincosamide and streptogramin antibiotics in Staphylococci isolated in Istanbul, Turkey

The purpose of this study was to investigate the prevalence and genetic mechanisms of erythromycin resistance in staphylococci. A total of 102 erythromycin resistant non-duplicate clinical isolates of staphylococci [78 coagulase negative stapylococci (CNS), 24 Staphylococcus aureus] were collected between October 2003 and August 2004 in Istanbul Faculty of Medicine in Turkey. The majority of the isolates were from blood and urine specimens. Antimicrobial susceptibilities were determined by the agar dilution procedure and the resistance phenotypes by the double disk induction test. A multiplex PCR was performed, using primers specific for erm(A), erm(B), erm(C), and msrA genes. Among the 78 CNS isolates, 57.8% expressed the MLSB-constitutive, 20.6% the MLSB-inducible, and 21.6% the MSB phenotypes. By PCR, 78.2% of these isolates harbored the erm(C) gene, 8.9% erm(A), 6.4% erm(B), and 11.5% msrA genes. In S. aureus, the constitutive MLSB (58.3%) was more common than the inducible phenotype (20.8%). erm(A) was detected in 50% and erm(C) in 62.5% of the isolates, while 37.5% contained both erm(A) and erm(Q. erm(C)-associated macrolide resistance was the most prevalent in CNS, while erm(C) and erm(A, Q was the most prevalent in S. aureus

Clonal distribution of vancomycin-resistantEnterococcus faeciumin Turkey and the new singleton ST733

Background The aim of this study was to provide information about the spread and characteristics of the vancomycin-resistantEnterococcus faeciumisolates (VREfm) in Turkey. Methods Seventy-one nonduplicate consecutive isolates of VREfm were obtained from various clinical specimens of inpatients treated at university or training hospitals in seven regions of Turkey. Further characteristics included antibiotic susceptibility testing, pulsed-field gel electrophoresis (PFGE) of SmaI-digested genomic DNA, and multilocus sequence typing (MLST) of selected isolates. The presence of vancomycin resistance and virulence genes (espandhyl) was investigated by polymerase chain reaction (PCR). Results All VREfm isolates had MICs to vancomycin of >= 32 mg/L and contained thevanA gene. The presence ofespgene was identified in 64 andhylin eight VREfm isolates. All VREfm showed the multiresistance phenotype, including ampicillin (99%), penicillin (99%), imipenem (99%), ciprofloxacin (87%), moxifloxacin (87%), erythromycin (97%), streptomycin (86%), gentamicin (82%), tetracycline (70%), and teicoplanin (99%). All were susceptible to tigecycline while quinupristin-dalfopristin (97%) and linezolid (93%) were the most active other agents. Analysis of the PFGE profiles showed that 53 (74.6%) VREfm isolates shared a similar electrophoretic profile, designed as type 1, and were closely related (>85%). The sequence type was identified by MLST in 44 VRE isolates with unrelated or closely related PFGE patterns. MLST revealed that nosocomial spread of VREfm resulted from dissemination of lineage C1E faeciumclones. Sequence types ST78, ST203, and ST117 were the most frequently isolated. This is the first report of ST733 around the world. Conclusions Lineage C1 clones are disseminated among clinical VREfm isolates in seven different regions in Turkey. Regarding VREfm isolates, the worldwide epidemic strains are in circulation in Turkey