Carbapenemase Gene Detection Protocol

Following the manufacturer's instructions, DNA of phenotypically confirmed XDR CP isolates were extracted using the Genomic DNA Purification Kit (Thermo Fisher Scientific, Waltham, MA, USA) and were used as templates for PCR using proper primers created by Macrogen® (Macrogen®, Madrid, Spain). The PCR amplification of the blaIMP, blaKPC, blaNDM, blaOXA-48, and blaVIM genes was performed using the annealing temperatures (Ta) and suitable primers as previously mentioned [18 (link), 25 (link)]. The amplified PCR results were examined using agarose gel electrophoresis, and using a 1000 bp DNA ladder (GeneRuler 1 kb, ThermoFisher Scientific, Waltham, MA, USA).