Carbapenemase investigation with rapid phenotypic test (RESIST-4 O.K.N.V) and comparison with PCR in carbapenem-resistant Enterobacterales strains

Melike  Yaşar-Duman; Feriha Çilli; Yamaç Tekintaş; Furkan Polat; Mine  Hoşgör-Limoncu

doi:10.18502/ijm.v14i3.9769

Melike Yaşar-Duman Department of Medical Microbiology, Faculty of Medicine, Ege University, Izmir, Turkey
Feriha Çilli Department of Medical Microbiology, Faculty of Medicine, Ege University, Izmir, Turkey
Yamaç Tekintaş Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Izmir Katip Celebi University, Izmir, Turkey
Furkan Polat Department of Medical Microbiology, Faculty of Medicine, Ege University, Izmir, Turkey
Mine Hoşgör-Limoncu Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Ege University, Izmir, Turkey

DOI: https://doi.org/10.18502/ijm.v14i3.9769

Keywords: Carbapenemases; Immunochromatographic test; Oxacillinase-48; Klebsiella pneumoniae

Abstract

Background and Objectives: RESİST-4 O.K.N.V. assay is a lateral immunochromatocraphic test for the identification of oxacillinase (OXA)-48-like, Klebsiella pneumoniae carbapenemase (KPC), New Delhi metallo-β-lactamase (NDM), and Verona integron-encoded metallo-β-lactamase (VIM) producing strains. It was aimed to evaluate the performance of the RESIST-4 O.K.N.V. test and to compare it with the reference method polymerase chain reaction (PCR). Also, the objective was to determine the distribution of carbapenemase types of CRE strains isolated in our hospital.

Materials and Methods: Between January 2016-October 2019, 187 strains isolated from clinical samples were included in this study. Bacterial identification was done using MALDI-TOF MS. Antibiotic susceptibility tests were studied with the VITEK-2 automated system. Meropenem minimum inhibitory concentrations (MICs) were determined by the gradi-ent test. All strains were studied with the RESIST-4 O.K.N.V. test and then the strains were selected for the PCR test. bla OXA-48 , bla NDM , bla KPC, and bla VIM were investigated with PCR. K. pneumoniae NCTC® 13438 (KWIKSTIKTM, Micro- biologics®,USA) was used as the positive control, E. coli ATTC® 25922 TM (Microbiologics®,USA) and three carbapen- em-sensitive clinical isolates were also used as the negative control.

Results: Meropenem MIC 50 and MIC 90 values were determined to be >32 mg/L. With PCR bla OXA-48 , bla NDM , bla KPC and bla VIM were detected in 79, 63, 20, and 4 strains, respectively. blaOXA-48 and bla NDM were found together in 51 of the iso- lates. bla OXA-48 , bla NDM , bla KPC, and bla VIM were not detected in two strains with carbapenem resistance in susceptibility tests. The sensitivity of the immunochromatographic test was 100% for OXA-48, KPC, and VIM but 84.1% for NDM. Specificity was determined as 100% for OXA-48, NDM, KPC, and VIM.

Conclusion: RESIST-4 O.K.N.V. test showed high sensitivity and specificity in detecting OXA-48, KPC, NDM, and VIM. type carbapenemases. However, it should be kept in mind that there may be false-negative results related to NDM.