EMERGENCE OF OXA-48 β-LACTAMASE IN

Dijana Varda-Brkić1123, Nataša Beader , Marko Čačić, Vranić-Ladavac 4 , Sanda Sardelić 567, Ines Jajić, Andrea Grisold , Gernot Zarfel7 , Josefa Luxner 7 , Zrinka Bošnjak 1,2 , Branka Bedenić 1,2 1University Hospital Center , Zagreb, Croatia,2 School of Medicine, University of Zagreb, Zagreb, Croatia,3 St. Antonius Hospital, Kleve, Germany,4 Public Health Institute of County, Croatia, 5University Hospital Center Split, Split, Croatia,6 Sisters of Mercy Hospital Center, Zagreb, Croatia,7 Institute for Microbiology, Hygiene and Environmental Medicine, University of Graz, Graz, Austria

Background Results Conclusions Previous studies found VIM-1, NDM-1 and The strains were uniformly resistant to The study demonstrated KPC-2 among CRE in Croatia with VIM-1 amoxicillin alone and combined with emergence of OXA-48β - being the most prevalent. Recently OXA-48 clavulanate, cefazoline, cefuroxime, lactamase in three hospital was identified in three hospital centers. ceftazidime, cefotaxime, ceftriaxone, centers located in different but uniformly susceptible to colistin geographic regions in Croatia and had variable resistance patterns to OXA-48 was first report in Turkey Material and methods carbapenems. Modified Hodge test in 2001 but later spread in was positive indicating the production Mediterranean and western Since 2012 surveillance system for CRE of carbapenemase.P henotypic testing European countries such as was implemented in University Hospital was positive for ESBL but negative for, Israel, France, Italy, Spain, Center (UHZ) in Croatia which inluded MBL, KPC and AmpC. Meropenem Germany, Switzerland, Belgium phenotypic and molecular identification of resistance was not transferred to E. and the Netherlands. carbapenemases inEnterobacteriaceae . In coli recipient strain. PCR revealed the total four carbapenem non-susceptible strains to be positive also for CTX-M- Most of the strains in western strains ofK. pneumoniae were found to be 15 and TEM-1. The strains from UHZ Europe were imported from positive for OXA-48 in three hospital coproduced OXA-48 and VIM-1. Two Turkey, Morocco, Egypt, Algeria or centers (University Hospital Center Zagreb- E. cloacae strains from UHZ belonged Lybia. UHZ, University Hospital Center Sisters of to the same ST-418. The patients did Mercy-UHM, University Hospital Split- not have a travel history to the UHS). The antimicrobial susceptibility was countries where OXA-48 is endemic. determined by broth microdilution method. Double-disk-synergy test (DDST) was performed to detect ESBLsand m odified Hodge test (MHT) was used to screen for production of carbapenemases. MBL E-test was used to screen for metallo-β- Table 1. Phenotypic tests, MICs, β-lactamase content, fluoroquinolone resistance genes, plasmid types and lactamases (MBLs). Additionally the rep-PCR clusters of carbapenem-resistant Enterobacteriaceae isolates were tested by combined disks test with EDTA or 3-aminophenylboronic to screen for KPC, MBLs or simoultaneous Protocol specimen department species date ESBL AmpC Hodge EDTA PBA BL qnr ST AMX AMC TZP CZ CAZ CXM CTX CRO FEP FOX IMI MEM ERT GM AMI CIP COL number production of MBL and KPC respectively. KBC- urine Outpatient E. cloacae 13. 12. +-++-VIM-1neg ST418l >128 >128 >128 >128 >128 >128 >128 >128 >128 >128 16 16 32 32 8 >128 0.12 209377 (previously 2013. OXA-48 The transferability of meropenem (281) hospitalized TEM-1 in UHZ) CTX-M-15 resistance was determined by conjugation KBC- urine Outpatient E. cloacae 17. 01. +-++-VIM-1neg ST418l >128 >128 >128 >128 >128 >128 >128 >128 >128 >128 16 16 32 32 16 16 0.25 - 8391(299) (previously 2014. OXA-48 employingE. coli A15R strain resistant to hospitalized TEM-1 in UHZ) CTX-M-15 sodium-azide. The presence of genes KBCSPLIT urine Dpt. Of K.pneumoniae 11.11. +----OXA-48neg ST37 >128 >128 >128 >128 >128 >128 >128 >128 >128 >128 32 32 32 32 16 16 0.12 UR17555 Medicine 2014. CTX-M15 encoding broad and extended-spectrum β- TEM-1 Sisters of urine urology K.pneumoniae 19.02. +-+--OXA-48neg NT >128 >128 >128 >128 >128 >128 >128 >128 >128 >128 1 0.5 16 >128 16 32 0.25 lactamases (blaSHV , bla TEM , bla CTX-M and Mercy-866 2015. CTX-M15 TEM-1 blaPER-1), plasmid-mediated AmpCβ - lactamases, group A carbapenemases (bla , bla , bla , bla ), metallo β- Abbreviations: AMX-amoxycillin; AMC-amoxycillin/clavulanate; CZ-cefazoline, CXM-cefuroxime; CAZ-ceftazidime; CRO-ceftriaxone; CTX-cefotaxime, FEP-cefepime; TZP-piperacillin/tazobactam; KPC SME IMI NDM FOX-cefoxitin; COL-colistin; IMP-imipenem; MEM-meropenem; ETP-ertapenem; GM-gentamicin; AMI-amikacin; CIP-ciprofloxacin; ESBL-combined disk test using cephalosporins alone and lactamases (blaVIM, bla IMP and bla NDM) , and combined with clavulanate for detection of ESBLs; AmpC- combined disk test using cephalosporins alone and combined with phenylboronic acid for detection of AmpC β-lactamases; PBA: carbapenem hydrolyzing oxacillinases combined disk test using carbapenems alone and combined with phenylboronic acid for detection of AmpC β-lactamases; EDTA: combined disk test using carbapenems alone and combined with

(blaOXA-48 ), was determined by PCR. phenylboronic acid for detection of MBLs; PBA-combined disk test using carbapenems alone and combined with phenylboronic acid for detection of KPCββ -lactamase; BL- -lactamase content; qnr-qnr genes- -encoding resistance to fluoroqinolones ;ST-sequence type ; NT-not tested