دراسة بعض جينات البايوفلم وجينات مقاومة الكوينولون في الإشريكية القولونية المسببة لألتهابات المسالك البولية المعزولة من مرضى التهاب المسالك البولية

Study of Some Biofilm Genes and Quinolone Resistance Genes in Uropathogenic Escherichia coli Isolated from Patients with Urinary tract infection

Abstract

Despite the health education and live style was better, some health problem still has the high rate of infection one of them was urinary tract infection (UTI). Morbidity and mortality associated with UTI were shown to be 150 million cases globally per year.
Urinary tract infection (UTI) is commonly caused by bacterial pathogens that often affect the urinary system including the kidney, ureters, bladder, and urethra. The bacterial pathogens invade the urinary tract involving the lower and the upper urinary tract and may sometimes spread to the bloodstream resulting in several clinical syndromes such as fever, flank pain, dysuria and hematuria.
This cross-sectional study aimed to Investigate of Some Biofilm Genes and Quinolone Resistance Genes in Uropathogenic Escherichia coli in UTI Patients included a total of (968) mid-stream urine samples collected from symptomatic patients of UTI of both sex with age range (1-80 years) were enrolled the two primary governmental hospitals in holy Karbala province, namely Imam Al-Hussein Medical City and Kerbala Pediatric Teaching Hospital During the period started from October 2023 to April 2024.
The sample collected was investigated for general urine examination and urine culture to identify E. coli bacteria and antibiotic susceptibility test using Vitek 2 compact system.
The positive bacterial growth from all samples was 261 (26.96 %) samples with 34 (3.51%) fungi and the remaining 673 (69.52%) being negative growth. After using Vitek 2 System implementation result, gram negative bacteria was the most common infectious agent which accounting for (64 %) of total positive culture and the E. coli was the highest percentage (61 %) of total positive bacterial growth and female patients was the higher infectious rate with this bacteria 77.45% in comparison to male 22.55%.
The antibiotic susceptibility test using Vitek 2 System show Ticarcillin was the highest resistant rate 88.89% followed by Piperacillin and Ampicillin while the most effective antibiotic was Imipenem and Meropenem with resistant rate about 6%.
One of the most important issues associated with the therapy of UTIs is the emergence and spread of multidrug-resistant, Topoisomerase IV which is composed of two subunits encoded by parC and par E, the primary function of which is chromosomal segregation following DNA replication, acquired quinolone resistance were attributable to mutations in quinolone resistance-determining region (QRDR) of topoisomerase IV. The Qnr proteins (qnrB and qnrS), which are members of the pentapeptide-repeat family, protect DNA gyrase and DNA topoisomerase IV from fluoroquinolone inhibition.
An important factor contributing to UPEC pathogenesis is the ability to form biofilms, CsgD consider as a major control and integration unit for biofilm formation due to its regulation of the expression of biofilm-associated extracellular matrix substances. While OmpT is an outer membrane serine protease functions as a defence mechanism for the bacteria by cleaving antimicrobial peptides secreted by the host epithelial cells. It also acts as a virulence factor by aiding in the spread of the bacteria within the host cells.
PCR revealed the presence of qnrB gene in (7.89 %) of the isolates, qnrS gene was in (18.4 %), ompT gene was in (68.4 %), parE gene was in 100% and csgD gene was in (97.4 %) among 38 E. coli isolates that showed highly antibiotic resistance specially to Fluoroquinolones group.
20 XDR E. coli isolates of 38 samples that involved in gene detection process, that showed the highest antibiotic resistance especially to Fluoroquinolones group, were sent to gene sequencing for both par E and csgD genes. The result of gene sequencing analysis of both par E gene and csgD genes, shows multiple variations were observed in different sites of each sample in both forward and reverse strand.
The genetic tree of the gene was analyzed using a specific program and it was compared with global isolates of E. coli bacteria.
The results of the phylogenetic tree of the gene par E (forward strand) for all 20 E. coli isolates when compared with the site NCBI of global isolates showed that the local isolates it occupied various positions within the phylogenetic tree, and did not lie at the same level with the location of NCBI strain except for the strain of sample ID number (23).
The results of the phylogenetic tree of the gene csgD (forward strand) for all 20 E. coli isolates when compared with the site NCBI of global isolates showed that the local isolates it occupied various positions within the phylogenetic tree, and did not lie at the same level with the location of NCBI strain except for the strain of sample ID number (37).
This study conclude that XDR E. coli was highly prevalent bacteria among UTI patients and that have highly mutation rate within the target-region that lead to make this bacteria more prevalent and more difficult to treat.