Efi Petinaki
University Hospital of Larissa, Greece
Title: Emergence of ST117 Enterococcus faecium carrying both vanA and vanB genes in Greece
Biography
Biography: Efi Petinaki
Abstract
Statement of the Problem: Vancomycin-resistant enterococci (VRE) are recognized as an important nosocomial problem worldwide, posing a particular threat in healthcare facilities (HCF). On November 2016, one E. faecium, carrier of both vanA/ vanB genes, was isolated from the blood cultures of a patient in the University Hospital of Larissa, Thessaly, Greece. The isolate, Efa-125, exhibited resistance to both vancomycin and teicoplanin (MIC vancomycin :256mg/L, MIC teicoplanin :256 mg/L), and was also resistant to various antimicrobial agents except, linezolid, daptomycin and tigecycline. Based on Multi Locus Sequence Typing (MLST), Efa-125 belonged to ST117 clone (Clonal Complex 17). Given that, few strains worldwide, were found to carry both vanA and vanB genes, the strain was further analyzed regarding the genetic units carrying vanA and vanB genes. Methodology: The genomic DNA of E. faecium Efa-125 was extracted using the DNA-Sorb-B kit (Sacace Biotechnologies S.r.l., Como, Italy). Plasmids and chromosomes were sequenced using the Illumina MiSeq platform (Illumina Inc., San Diego, CA, USA). Initial paired-end reads were quality trimmed using Trimmomatic tool and assembled via de Bruijn graph-based de novo assembler SPAdes. The sequence gaps were filled by a PCR-based strategy and Sanger sequencing. For sequence analysis and annotation, the BLAST algorithm (www.ncbi.nlm.nih.gov/BLAST), the ISFinder database (www-is.biotoul.fr/), and open reading frame (ORF) finder tool (www.bioinformatics.org/sms/) were utilized. The nucleotide sequence of the plasmid pEfa-125gr has been assigned GenBank accession number KY320277.Findings: The vanA gene was carried on a plasmid (29,320 bp) exhibiting high similarity to pA6981, previously characterized from E. gallinarum A6981, whereas vanB was part of a Tn1549-like transposon integrated into its chromosome.Conclusion & Significance: In Greece is the first detection of a vanA-vanB genotype/VanA phenotype E. faecium caused bacterieamia. Until now, vanA/vanB E. faecium strains are isolated in Middle East and express a VanB phenotype. This report indicates an evolving VRE epidemiology.