MOLECULAR DETECTION OF VIRULENCE GENES IN KLEBSIELLA PNEUMONIAE ISOLATES FROM WASIT PROVINCE, IRAQ

Abstract

The most common kind of nosocomial infection is an infection of the urinary tract (UTI), and K. pneumonia is the second most common Gram-negative bacterial cause of UTIs. The rise of bacterial pathogens that are resistant to several drugs is a serious global health concern. Overuse of antibiotics has reduced our treatment choices for K. pneumoniae and made appropriate management of this bacterial illness more challenging. ‎

The ability to produce biofilm was examined by using microtiter plate ‎‎(MTP) and modified Congo red agar (MCRA), in MCRA method, among ‎the 40 clinical strains 35(87.5%) were biofilm producers.33 (82.5%) ‎were strong biofilm-producers with black color, 2(5%) were classified as ‎weak biofilm-producer in orang(grey) colony, whilst 5 (12.5%) were ‎pink isolates of Klebsiella pneumonia none producers biofilm. In MTP ‎method , the 40 K. pneumonia isolates tested, there were 36 (90%) ‎isolates as biofilm producer and 4 (10%) isolates that were not biofilm ‎producers. Among biofilm producers, there were 32 (80%) isolates as ‎strong, 2 (5%) isolates as moderate, and 2 (5%) isolates identified as ‎weak biofilm producers.‎

16S rRNA confirms the molecular identification of k pneumonia isolates. Utilizing the NanoDrop instrument, the concentration and purity were determined. Purity was between 1.6 and 2.0, while concentration ranged from 50 to 360 ng/l. The fimH gene, with 550 bp was found in 82% of the isolates, 226 bp mrkD gene gave 70% positive results. ‎

The mrkD primer nucleotide sequences of six samples were analyzed and compared to the NCBI database. Three samples had a perfect match, as determined by the nucleotide sequence analysis, while the remaining samples had a match rate of between 97 and 99 percent. Transition A/G, Transition T/C, Transversion A/C, and Gaps were among the several genetic variations found. ‎