Sample collection and microbial detection

Urine specimens were cultured on blood agar and MacConkey agar plates (Hi-Media, India) at 37°C for 24 h. The grown isolates were recognized by the morphology of colonies, Gram-staining, and biochemical tests properties. K. pneumoniae colonies on blood agar medium were large, cupola shape, and mucoid that move to merge. On MacConkey medium, K. pneumoniae colonies were large, mucoid dark pink, which specifies the lactose fermentation. The biochemical tests for K. pneumoniae included of negative indole, positive urease, positive VP, positive Simmons’ citrate agar, variable magnetic resonance, and no motile. Bacterial isolates were refrigerated on nutrient agar plates or frozen (−70°C) in microtubes containing 15% glycerol and tryptic soy broth medium.[2] Afterward, molecular confirmation was performed by PCR of the ureD gene. Analysis for the presence of the ureD gene demonstrated that all the isolates confirmed as K. pneumoniae were positive for the ureD gene.