Analysis performed by the NRC

 


  • Confirmation of bacterial identification to species level using MALDI-TOF MS

  • In vitro susceptibility testing to antimicrobial drugs by disc diffusion according to CLSI guidelines
  • Determination of MICs by broth microdilution method according the CLSI guidelines and interpretative breakpoints

  • Carbapenem hydrolysis-based tests
  • Inhibitors-based combination disk tests:
    • For the detection of ESBL and/or hyperproduced AmpC
    • For the detection of carbapenemase
  • Extended susceptibility disk diffusion testing against aminoglycosides for the detection of 16S rRNA methylases

  • Loop mediated isothermal amplification (LAMP) targeting NDM, VIM, OXA-48, OXA-181, KPC and CTX-M group 1 and 9
  • Immunochromatographic detection of carbapenemase (OXA-48 and KPC)
  • End-point PCR simplex or multiplex targeting
    • carbapenemases (NDM, VIM, KPC, IMP, OXA-48, OXA-427, BES, IMI, SIM, GIM, DIM, FIM, SMB, SME, SPM)  
    • minor ESBL (GES, VEB, PER and BEL)
    • oxacillinases groups (OXA-1,OXA-2,OXA-9,OXA-10, OXA-18,OXA-20 and OXA-198)
    • ESBL (TEM, SHV, CTX-M group 1,  2 and 9 )
    • plasmidic cephalosporinases (MOX, CMY, DHA, ACC, ACT/MIR, FOX)
    • 16S rRNA methylases (ArmA, Rmt A-B-C-D-E-F-G, NpmA)
    • plasmid-borne quinolone resistance genes (QnrA, QnrB, QnrS)
  • DNA microarray
    • Check-MDR assays targeting ESBLs, carbapenemases and plasmidic cephalosporinase genes
  • Other genetic characterization methods:
    • Sequencing of resistance genes  and the genetic environment
    • Plasmid extraction by Kieser method
    • Conjugation, transformation and cloning
    • PCR-based replicon typing : method based on replicons (Inc/rep PCR) of the major plasmid incompatibility groups (Caratolli A. et al , JAC. 2010 Dec;65(12):2518-29)
    • Multiplex End-point PCR detecting capsular types (K1, K2, K5, K54 and K57) and virulence factors (rmpA,  wcaG, terW, iutA and silS) in K.pneumoniae
    • Quadruplex End-point PCR assigning E.coli to the following phylo-groups (A,B1,B2,C,D,E and F)

Semi-automated rep-PCR

Pulse-Field Gel Electrophoresis https://nrchm.wiv-isp.be/fr/centres_ref_labo/esbl_carbapenemase_prod_enterobacteriaceae/default.aspx

MLST typing  : PCR and sequencing of 7 house keeping genes and definition of ST according the website

E coli : http://mlst.ucc.ie/mlst/dbs/ Ecoli

K.pneumoniae: http://www.pasteur.fr/recherche/genopole/PF8/mlst

Rue Docteur Gaston Therasse 1, 5530 Yvoir (Belgique)