Application of Molecular Methods for Carbapenemase Detection.

This study has evaluated the correlation between different carbapenemases detection methods on carbapenem non-susceptible Klebsiella pneumoniae strains from Northern and Eastern Europe; 31 institutions in 9 countries participated in the research project, namely Finland, Estonia, Latvia, Lithuania, Russia, St. Petersburg, Poland, Belarus, Ukraine, and Georgia. During the research program, a total of 5,001 clinical K. pneumoniae isolates were screened for any carbapenem non-susceptibility by the disk diffusion method, Vitek 2 or Phoenix system following the EUCAST guideline on detection of resistance mechanisms, version 1.0. Strains isolated from outpatients and hospitalized patients from April 2015 to June 2015 were included. All types of samples (blood, pus, urine, etc.) excluding fecal screening or fecal colonization samples have been represented. In total, 171 carbapenemase screening-positive K. pneumoniae isolates (3.42%) were found and characterized. Several methods were used for detection of carbapenemases production, including Luminex assay (PCR and hybridization), whole genome sequencing, MALDI-TOF based Imipenem degradation assay, and immunochromatography testing. Minimal inhibitory concentration determination for Meropenem by agar-based gradient method was also used. Finally, 83 K. pneumoniae strains were carbapenemase negative by all confirmation methods (49.4% of all screening-positive ones), 74 - positive by three methods (44.0%), 8 - positive by two methods (4.8%) and 3 - positive by only one method (1.8%). The sensitivity of the tests was 96.3% for Whole genome sequencing and MALDI-TOF assay (both three undetected cases), and 95.1% for Luminex-Carba (4 undetected cases). The most commonly detected carbapenemases were NDM (n = 54) and OXA-48 (n = 26), followed by KPC-2, VIM-5, and OXA-72 (one case of each). Our results showed that different types of carbapenemases can be detected in the countries involved in the project. The sensitivity of our methods for carbapenemase detection (including screening as a first step and further confirmation tests) was >95%, but we would recommend using different methods to increase the sensitivity of detection and make it more precise.

Antimicrobial resistance of Escherichia coli isolates from outpatient urinary tract infections in women in six European countries including Russia.

OBJECTIVES: In the Northern Dimension Antibiotic Resistance Study (NoDARS), Finland, Germany, Latvia, Poland, Russia and Sweden collected urine samples from outpatient women (aged 18-65years) with symptoms of uncomplicated urinary tract infection (UTI) to investigate the levels of antimicrobial resistance (AMR) among Escherichia coli isolates. METHODS: A total of 775 E. coli isolates from 1280 clinical urine samples were collected from October 2015 to January 2017. Antimicrobial susceptibility testing was performed and the results were interpreted according to European Committee on Antimicrobial Susceptibility Testing (EUCAST) criteria. RESULTS: Overall AMR rates to the commonly used antibiotics nitrofurantoin, fosfomycin and mecillinam (except for Germany that was missing a result for mecillinam) were 1.2%, 1.3% and 4.1%, respectively. The highest overall resistance rates were determined for ampicillin (39.6%), trimethoprim (23.8%), trimethoprim/sulfamethoxazole (22.4%), amoxicillin/clavulanic acid (16.7%) and ciprofloxacin (15.1%), varying significantly between countries. The rate of extended-spectrum ß-lactamase (ESBL) production was 8.7%. None of the isolates showed resistance to meropenem. CONCLUSIONS: In most cases, low AMR rates were detected against the first-line antibiotics recommended in national UTI treatment guidelines, giving support to their future use. These results also support the European Association of Urology guidelines stating that nitrofurantoin, fosfomycin and mecillinam are viable treatment options for uncomplicated UTI.

Dissemination of a carbapenem-resistant Acinetobacter baumannii strain belonging to international clone II/sequence type 2 and harboring a novel AbaR4-like resistance island in Latvia.

An outbreak of hospital-acquired Acinetobacter baumannii infections, caused by a bla(OXA-23)-positive carbapenem-resistant strain belonging to international clone II/ST2, was detected in Latvia. The strain was partially equipped with the armA gene and the intI1-aacA4-catB8-aadA1-qacEΔ1 class 1 integron. In addition, the strain carried AbaR25, a novel AbaR4-like resistance island of ~46,500 bp containing structures similar to the previously described AbaR22 and Tn6167 islands. AbaR25 was characterized by the occurrence of a second copy of Tn6022a interrupted by Tn2006 carrying the bla(OXA-23) gene.

Outbreak of CTX-M-15-producing Klebsiella pneumoniae of sequence type 199 in a Latvian teaching hospital.

Previous studies on the epidemiology of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae in Latvia are lacking. ESBL-producing Klebsiella pneumoniae (n = 32) were subjected to pulsed-field gel electrophoresis (PFGE) and selected isolates to multi-locus sequence typing (MLST). Species identification and susceptibility testing were performed using VITEK2, and sequencing of bla(CTX-M) was performed in selected isolates. PFGE revealed one major clone (n = 23), with most of the isolates derived from the ICU. The clone harboured bla(CTX-M-15), was sequence type 199 and comprised two ertapenem non-susceptible isolates. This is the first report of an ESBL outbreak in Latvia, and calls for increased epidemiological typing of ESBL-producing Enterobacteriaceae, as well as improved infection control routines.