This cross-sectional study collected 100 clinical specimens from Hilla Teaching Hospital and Marjan Medical City patients aged 3 to 55, 65% of which were male and 35% female. Forty strains of K. pneumonia were isolated from 100 specimens from various infection locations by morphological, microscopical, biochemical, and Vitek II system testing. Eleven (27.5%) of the forty isolates were from sputum, nine (20%) from urine, seven (17.5%) from wound swabs, six (15%) from burn swabs, five (12.5%) from burn tissue, and three (7.5%) from ear swabs. K. pneumonia was isolated from 26 men (65%) and 14 women (35%). All of the possible K. pneumonia isolates had their DNA taken and ran through a standard PCR for pilv-l gene primer amplification using the sequences, the results revealed that only 16(40%) of the 40 K. pneumonia isolates were related to K. pneumonia258 by sharing the same 320 bp DNA fragment with the allelic ladder. Some virulence genes were detected in 16 K. pneumoniae258 isolates. The results showed that FimH all 16(100%) were positive to FimH gene of K. pneumoniae258 isolates at (688bp). However, mrkD gene were detected 13/16(81.2%) were positive to mrkD gene of K. pneumoniae258 isolates at (240bp). Objective: The aim of present study is to isolation of the K. pneumoniae 258 and the detection of K. pneumoniae 258 to some virulence genes.