Evaluation Of The Two Commercial Methods Based On Broth Microdilution Against Reference Microdilution Method For Klebsiella Pneumoniae Isolates

A rapid and reliable method for antimicrobial susceptibility test of colistin is needed because of increasing numbers of multi-resistant gram negative bacterial infections and simultaneus increasing of colistin resistance. Although broth microdilution (BMD) is recommended by the Clinical and Laboratory Standards Institute (CLSI) and the European Committee for Antimicrobial Susceptibility Testing (EUCAST) as a reference method, the use in routine laboratory practice is limited because of the difficulties in application and time-consuming characteristics. Recently, many BMD based commercial products were developed. The study was aimed to compare the results of the two commercial systems available for the detection of colistin sensitivity with the reference method. Totally 38 Klebsiella pneumoniae strains isolated from various clinical specimens between 2017-2018 were included in our study. Identification and antibiotic susceptibility tests were performed with "matrix-assisted laser desorption/ionization time-of-flight, mass spectrometry (MALDI-TOF MS)" and Vitek-2 (bioMerieux, Marcyl'Etoile, France) systems. BMD tests were performed with Sensititre (Sensititre custom plate, Thermo Fisher Scientific, UK) and Micronaut MIC-Strip (Merlin Diagnostika GmbH, Germany) kits. Commercial BMD assays containing dehydrated colistin in the concentration range of 0.0625-128 mg/L were tested with 0.5 McFarland bacterial suspensions prepared according to the manufacturer recommendations. The reference BMD test was performed by following the recommendations of the International Organization for Standardization (ISO-20776-1). ATCC 25922 colistin-susceptible Escherichia coil and NCTC 13846 (mcr-1 positive) colistin-resistant E.coll strains were used as the quality control strains. According to the recommendations of EUCAST version 9.0, strains with minimum inhibitory concentration value of <= 2 mg/L were considered susceptible and strains > 2 mg/L as resistant. Thirty-five isolates were resistant to colistin by the reference method and three of them were susceptible. The Sensititre kit detected a very major error (2.8%) in one isolate; no major or very major errors were detected for the Micronaut kit. The essential and categorical agreement of the Sensititre and Micronaut kits with the reference method was defined as 74-76% and 97-100%, respectively. Colistin is the last agent for the treatment of the multi drug resistant severe bacterial infections so major and very major error for colistin should be considered equally serious. Although a very major error was detected by the Sensititre kit in one isolate, the categorical agreement of both commercial kits was greater than 90% when compared with the reference method. It was concluded that, commercially available, BMD based systems that do not require additional equipment and experience can be routinely used.