title: Detection of OXA-48-type carbapenemase-producing Enterobacteriaceae in diagnostic laboratories can be enhanced by addition of bicarbonates to cultivation media or reaction buffers.
abstract: Carbapenemase-mediated resistance to carbapenems in Enterobacteriaceae has become the main challenge in the treatment and prevention of infections recently. The partially unnoticed spread of OXA-48-type carbapenemase producers is usually assigned to low minimum inhibitory concentrations (MICs) of carbapenems that OXA-48-producing isolates often display. Therefore, there is an urgent need of specific and sensitive methods for isolation and detection of OXA-48 producers in clinical microbiology diagnostics. The influence of bicarbonates on carbapenem MICs against carbapenemase-producing Enterobacteriaceae was tested. We also checked whether the addition of bicarbonates to liquid media supplemented with meropenem may facilitate the selective enrichment of various carbapenemase producers in cultures. Furthermore, the sensitivity of carbapenemase confirmation by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS) and spectrophotometric hydrolysis assays upon the addition of NH4HCO3 was examined. The addition of NaHCO3 significantly increased MICs of ertapenem and meropenem for OXA-48 producers. Furthermore, liquid media supplemented with NaHCO3 and meropenem were reliable for the selective enrichment of carbapenemase producers. The presence of NH4HCO3 in buffers used in the spectrophotometric and MALDI-TOF MS carbapenemase detection increased the sensitivity of that assay. Our results demonstrate that bicarbonates in media or reaction buffers can enhance the sensitivity of screening methods and diagnostic tests for carbapenemase producers.
authors.affiliation: Department of Microbiology, Faculty of Medicine and University Hospital in Plzen, Charles University in Prague, Alej Svobody 80, 304 60, Plzen, Czech Republic.
authors.forename: Vendula
authors.initials: V
authors.lastname: Studentova
authors.forename: Costas C
authors.initials: CC
authors.lastname: Papagiannitsis
authors.forename: Radoslaw
authors.initials: R
authors.lastname: Izdebski
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Pfeifer
authors.forename: Eva
authors.initials: E
authors.lastname: Chudackova
authors.forename: Tamara
authors.initials: T
authors.lastname: Bergerova
authors.forename: Marek
authors.initials: M
authors.lastname: Gniadkowski
authors.forename: Jaroslav
authors.initials: J
authors.lastname: Hrabak
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis / Q000378:metabolism*; D001639:Bicarbonates; D002021:Buffers; D003470:Culture Media / Q000737:chemistry*; D004755:Enterobacteriaceae / Q000201:enzymology* / Q000302:isolation & purification*; D004756:Enterobacteriaceae Infections / Q000175:diagnosis / Q000382:microbiology*; D008826:Microbial Sensitivity Tests / Q000379:methods*; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D001618:beta-Lactamases / Q000032:analysis / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D001639:Bicarbonates; D002021:Buffers; D003470:Culture Media; D001618:beta-Lactamases; C063912:carbapenemase