Study of virulence genes variation of uropathogenic E. coli using of multilocus sequence typing technique in patients of kidney failure in Anbar province

Around 120 clinical specimens in the current work were collected from male and female of Iraqi patients who presented Ramadi teaching hospital in the Al-Anbar governorate. The distribution of these specimens was as follows: 60 urine specimens were collected with urinary tract infections, and extra 60 urine samples were taken from patients with renal failure (UTIs). All patient urine specimens were cultured on MacConkey agar, blood agar, and Eosin Methylene blue. All patient specimens were culture-positive. According to the cultural, morphological, biochemical characteristics, and VITEK the bacterial strains were identified. Each isolate of E. coli was subjected to bacterial DNA extraction in order to identify five important genes: 16S RNA, FimH, Hly, Cnf, and PapC. These genes were amplified using the Polymerase Chain Reaction (PCR) technique. In the current study, molecular typing for bacterial genes was studied using multilocus sequence typing (MLST). Concerning renal failure patients, only 27 (45%) of the samples were identified as E. coli, while the remaining 33 (55%) were identified as other species. Likewise, among UTI patients, only 43 (71.7%) were identified as E. coli, and the remaining percentage, 17 (28.3%), were identified as other species. The clinical specimens of renal failure and UTI among patients were distributed according to gender, into 33 (55%) and 22 (36.6%) in males and 27 (45%) and 38 (63.4%) in females, respectively. According to the findings of the current study, male and female patients with renal failure between the ages of 50 and 60 had the highest percentage rate (29.9% and 25.9%, respectively), followed by those between the ages of 40 and 50 (24.2% and 25.9% respectively). Concerning UTIs patients in the present work study, male patients with UTI were more likely to develop it between the ages of 20 and 30 and 30 and 40 years (22.7 percent in both age groups). In addition, the results showed that among females, the age group 30–40 had the highest rate of UTIs (28.9%), followed by the 40–50 age group (23.60%). The bacterial isolates which isolated from UTI patients were recorded highly resistance for set of antibiotics under study as follow: only 42(97.67%), 41(95.35%), 40(93.02%), 40 (93.02%), 40 (93.02%), 39 (90.70%) and 38 (88.37%) for Cefepime, piperacillin, trimethoprim/sulfamethoxazole, ticarcillin/clavulanic acid, ceftazidime, ticarcillin, ciprofloxacin, and aztreonam respectively. Likewise, the isolates were recorded highly sensitivity for set of antibiotics which as follow: only 36 (83.72%), 37 (86.05%), 37(86.05%), 38(90.70%) 39(90.70%) and 41(95.35%) for ertapenem, levofloxacin, imipenem, meropenem, tigecycline, and amikacin respectively. Moreover, the resistance strains that isolated from patients with renal failure for antibiotics under study were noted highly percentage for cefazolin 21 (77.78%), ceftazidime 22 (81.48%), and ciprofloxacin 23(85.19%). In addition, all strains which isolated from renal failure patients were recorded highly sensitivity for tobramycin 22(81.48%), tigecycline 26(96.30%), levofloxacin 27(100%), and amikacin 27 (100%) respectively. Concerning virulence genes FimH, PapC, Cnf and Hly, all strains were contained these genes as well as 16S RNA where all isolates were reported 100% for each genes 16S RNA, FimH, PapC, Cnf and Hly.