Candida auris has recently emerged as a multidrug-resistant yeast implicated in various healthcare-associated invasive infections and hospital outbreaks. In the current study, we report the first five intensive care unit (ICU) cases affected by C. auris isolates in Greece, during October 2020–January 2022. The ICU of the hospital was converted to a COVID-19 unit on 25 February 2021, during the third wave of COVID-19 in Greece. Identification of the isolates was confirmed by Matrix Assisted Laser Desorption Ionization Time of Flight mass spectroscopy (MALDI-TOF]. Antifungal susceptibility testing was performed by the EUCAST broth microdilution method. Based on the tentative CDC MIC breakpoints, all five C. auris isolates were resistant to fluconazole (≥32 μg/mL), while three of them exhibited resistance to amphotericin B (≥2 μg/mL). The environmental screening also revealed the dissemination of C. auris in the ICU. Molecular characterization of C. auris clinical and environmental isolates was performed by MultiLocus Sequence Typing (MLST) of a set of four genetic loci, namely ITS, D1/D2, RPB1 and RPB2, encoding for the internal transcribed spacer region (ITS) of the ribosomal subunit, the large ribosomal subunit region and the RNA polymerase II largest subunit, respectively. MLST analysis showed that all isolates possessed identical sequences in the four genetic loci and clustered with the South Asian clade I strains. Additionally, PCR amplification and sequencing of the CJJ09_001802 genetic locus, encoding for the “nucleolar protein 58” that contains clade-specific repeats was performed. Sanger sequence analysis of the TCCTTCTTC repeats within CJJ09_001802 locus also assigned the C. auris isolates to the South Asian clade I. Our study confirms that C. auris is an emerging yeast pathogen in our region, especially in the setting of the ongoing COVID-19 worldwide pandemic. Adherence to strict infection control is needed to restrain further spread of the pathogen.
An Escherichia coli clinical strain resistant to all β-lactams except carbapenems was isolated in a Greek hospital. Analysis of β-lactamase content by isoelectric focusing, PCR assays specific for various bla genes, and DNA sequencing showed that the strain produced TEM-1, a Citrobacter freundii AmpC-related cephalosporinase, and CTX-M-3. The blaCTX-M-3 gene was carried by a 120-kb plasmid that was readily transferable to a susceptible E. coli host.
Objectives: Streptococcus pyogenes causes superficial infections, but can also cause deep-seated infections and toxin-mediated diseases. In the present study, phylogenetic and in silico prediction analyses were performed on an antimicrobial-resistant M1UK S. pyogenes strain causing severe clinical manifestations during the current surge of invasive group A Streptococcus (iGAS) disease. Patient and Methods: A 40-year-old patient was admitted to the hospital with fever, chest pain, and fatigue. Based on the clinical and laboratory findings a diagnosis of sepsis with disseminated intravascular coagulation, community-acquired pneumonia, pleural empyema and streptococcal toxic shock syndrome was made. Microbial identification was performed by multiplex PCR and conventional culturing. Furthermore, antimicrobial susceptibility testing, whole genome sequencing, phylogenomic analysis, and in silico prediction analysis of antimicrobial resistance genes and virulence factors were performed. Results: S. pyogenes isolates were detected in pleural fluid and sputum of the patient. Both isolates belonged to the M1UK lineage of the emm1/ST28 clone, being closely related with an M1UK GAS strain from Australia. They exhibited resistance to erythromycin and clindamycin, and susceptibility-increased exposure to levofloxacin and carried genes encoding for protein homologues of antibiotic efflux pumps. Moreover, several virulence factors, and a previously described single-nucleotide polymorphism in the 5′ transcriptional leader sequence of the ssrA gene, which enhances expression of SpeA, were detected. Conclusions: The present antimicrobial-resistant M1UK S. pyogenes strain represents the first report of this emerging lineage associated with such manifestations of iGAS disease.
The aim of this study was to determine the rate and the mutations of genes involved to the first-line antituberculous drugs' resistance of M. tuberculosis/canettii isolated in Central Greece from 2010 to 2019. During the study period, the rate of resistance to isoniazid, rifampicin, ethambutol, and pyrazinamide was 5.4%, 0.4%, 1.1%, and 1.1%, respectively. All phenotypically resistant isolates (14 to isoniazid, 3 to ethambutol, 3 to pyrazinamide, and 1 to rifampicin) and 17 susceptible isolates (control group) were tested for the presence of mutations/alterations/polymorphisms by PCR followed by sequencing analysis. The molecular typing of isolates was based on multispacer sequence typing. Despite the phenotypic resistance, mutations were detected in 13 of 21 isolates (11 isoniazid resistant, 1 rifampicin, and 1 pyrazinamide resistant). Four isoniazid-resistant strains carried the most common mutations S315T and C-15T, whereas the remaining seven isolates carried either less known (E399, A162, W477STOP, S94A, G-48A, C-54T, C-17T, L203, A196, S124, and K367) or novel (D74N, G691S, Ains-85, and D171G); none of the susceptible strains was found to be positive for any novel mutation. The two single rifampicin- and pyrazinamide-resistant strains carried the known mutations S450L (also referred as S531L) and L182W, respectively. The presence of uncommon or novel mutations conferring resistance to isoniazid (INH) creates a diagnostic problem in the routine microbiological laboratory, since commercial methods are focused on the detection of the most common mechanisms of resistance (S315T, C-15T, A-16G, T-8C, and T-8A), therefore, fail to detect such strains. The regional differences in the frequencies of mutations associated with resistance to the first-line drugs provide hints for the development of better molecular-based diagnostic tests.
During 2014-2016, a total of 248 carbapenem-resistant Klebsiella pneumoniae (CARB-R Kp) were recovered in a Greek intensive care unit (ICU), the colistin resistance (COL-R) rates among CARB-R Kp from bloodstream infections (BSIs) were determined, and molecular characterization and the in vitro susceptibility of CARB-R+COL-R Kp to ceftazidime/avibactam were performed. The majority of CARB-R Kp from BSIs (n = 53) were OXA-48 (43.4%) and KPC (33.9%) producers, but no statistically significant differences were observed for the clinical characteristics of ICU patients affected by OXA-48 and other carbapenemase-producing K. pneumoniae. CARB-R+COL-R Kp (n = 28) represented 52.8% of 53 CARB-R Kp recovered from BSIs. The increase in the COL-R rates from 2014 to 2015 was mainly associated with the diffusion of extensively drug-resistant (XDR) OXA-48-co-producing CTX-M-15-like K. pneumoniae, assigned to multilocus-sequence typing ST101, possessing alterations in the mgrB loci. Ceftazidime/avibactam was active against all OXA-48 and KPC producers. Thus, the spread of XDR Kp possessing different types of carbapenemases further complicates the infection control strategies for the management of XDR Kp, whereas ceftazidime/avibactam may be a reasonable alternative to colistin for the treatment of XDR Kp in settings with low prevalence of metallo-β lactamase-producing K. pneumoniae.
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