Treatment of pandrug-resistant isolates often necessitates combination therapy. Checkerboard synergy and time-killing assay tests were performed to evaluate the benefits of a triple combination with meropenem, ertapenem, and colistin against 10 colistin-resistant K. pneumoniae clinical isolates harboring different β-lactamases. (blaOXA-48, blaNDM). In this study, ertapenem and meropenem (ERT/MEM), meropenem and colistin (MEM/COL), ertapenem, meropenem and colistin (ERT/MEM/COL) combinations were tested using checkerboard techniques and time-kill assays of each antibiotic alone and in combination against 10 colistin-resistant clinical K. pneumoniae isolates. An analysis of K. pneumoniae isolate B6 using a scanning electron microscope revealed morphologic changes in the cell surface after treatment with each antibiotic both alone and in combination. The whole genome of K. pneumoniae KPNB1 was sequenced using an Ion Torrent PGM sequencer. According to the checkboard results, synergistic combinations were observed with ertapenem/meropenem (5/10 isolates), meropenem/colistin (7/10) and ertapenem/meropenem/colistin (9/10); no antagonism was observed for all combinations. For the time-kill assay results; synergism and bactericidal effects were observed with meropenem/colistin (10/10) and with ertapenem/meropenem/colistin (10/10) combinations, and an indifference effect was observed with the ertapenem and meropenem (10/10) combination. Strain number 1 was found 100% identical to Klebsiella pneumoniae subsp. pneumoniae HS11286 according to the outcomes of complete genome sequence analysis, and the strain carried the genes blaOXA-181, blaCTXM-15, blaNDM, arr-3, aac (6′)-Ib-cr, rmtF, and catB1. Using double carbapenem antibiotics with colistin could be a potential alternative to treat colistin and carbapenem-resistant K. pneumoniae. The present study is the first Turkish report of OXA–181-type carbapenemase causing colistin resistance.
6 October 2020 in Journal of Clinical Laboratory Analysis
Background: The aim of this study was to provide information about the spread and characteristics of the vancomycin‐resistant Enterococcus faecium isolates (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 (esp and hyl) was investigated by polymerase chain reaction (PCR).Results: All VREfm isolates had MICs to vancomycin of ≥32 mg/L and contained the vanA gene. The presence of esp gene was identified in 64 and hyl in 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 C1 E faecium clones. 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.
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 β-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.
Background: In the post-pandemic era, respiratory viruses continue to cause substantial morbidity and mortality among hospitalized adults. SARS-CoV-2 and influenza remain the most common pathogens, while RSV and rhinovirus have re-emerged as relevant causes of severe illness. This study compared the characteristics and outcomes of virus-specific infections detected by multiplex real-time PCR over two consecutive seasons. Methods: This retrospective cohort study was conducted at a 1010-bed tertiary-care hospital in Türkiye between June 2022 and June 2024. Adults hospitalized with at least one respiratory virus detected by MRT-PCR were included. Demographic, clinical, and laboratory data were analyzed. Pathogen-specific comparisons were limited to monoinfections, and predictors of in-hospital mortality were identified using multivariable logistic regression. Results: Among 518 admissions, influenza (33.6%) and SARS-CoV-2 (29.3%) were the predominant pathogens, followed by rhinovirus (11.2%), RSV (6.6%), and other respiratory viruses (19.6%). Overall in-hospital mortality was 26.6%. Mortality differed across virus groups in unadjusted analyses, being highest in SARS-CoV-2 and RSV and lowest in rhinovirus. Non-survivors were older, more comorbid, more often immunosuppressed, and more likely to require oxygen therapy or ICU care at sampling. In multivariable analysis, independent predictors of mortality were ICU location at sampling (aOR 5.52), oxygen requirement (aOR 3.39), immunosuppression (aOR 3.67), older age (per 10-year increase: aOR 1.25), and secondary bacterial infection (aOR 7.00). Viral etiology, including SARS-CoV-2, was not independently associated with mortality after adjustment. Conclusions: Among hospitalized adults, mortality was driven primarily by host-related factors and secondary bacterial infection rather than by viral etiology. These findings highlight the need for strengthened adult immunization programs, reliable respiratory virus surveillance, the prevention of bacterial superinfection, and the development of and equitable access to effective vaccines and antiviral therapies to reduce severe outcomes in high-risk adults.
Rapid, simple, and accurate laboratory detection of carbapenemases is very important for proper antibiotic therapy and infection control. In this study, carbapenem-nonsusceptible Enterobacteriaceae (CRE) isolates were used to evaluate the performance of a new lateral flow immunochromatographic (IC) assay, the OXA-48 and KPC K-SeT assay, and modified Blue-Carba test (BCT) for the rapid detection of OXA-48 carbapenemase in comparison with polymerase chain reaction (PCR) amplification. These CREs of various enterobacterial species were isolated from various clinical samples including OXA-48 (47), NDM-1 (6), KPC-1 (1), IMP-1 (1), VIM-2,-4 (2), IMP-2 (1), OXA-51 (1), and OXA-23 (1) producers. The OXA-48 K-SeT test detected all OXA-48 carbapenemase producers with 100% sensitivity and specificity. The BCT detected carbapenemase producers with 93% sensitivity and 100% specificity. Both IC assays and BCT tests have good performance and are easy to perform, rapid, simple to interpret, and highly sensitive. We suggest that BCT can be used initially as an accurate, inexpensive, and rapid phenotypic confirmation test to identify Class A, B, and D carbapenemases in the routine diagnostic microbiology laboratory, thus allowing the detection of carbapenemase activity directly from bacterial cultures.
16 September 2022 in Acta Microbiologica Et Immunologica Hungarica
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-meropenem (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 indifferent 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.
About Me
Associate profesor at Eskisehir Osmangazi Univeristy, Faculty of Medicine, Department of Medical Microbiology
Associate profesor at Eskisehir Osmangazi Univeristy, Faculty of Medicine, Department of Medical Microbiology