دور البروتين المرتبط بالهيبارين وجينات الضراوة في البكتريا المنتجة لإنزيمات بيتا لاكتاماز واسعة الطيف بين مرضى عدوى المسالك البولية

The Role of Heparin Binding Protein and Virulence Genes of ESBL Uropathogenic Bacteria in Patients With Urinary Tract Infection

Abstract

Extended-Spectrum Beta-Lactamase (ESBL) producing bacteria are a group of bacteria that have developed resistance to a wide range of commonly used antibiotics, including many penicillins and cephalosporins. The presence of the CNF, A/E, BLA OXA-48, BLA CTX-M, and heparin binding protein in the urine of Escherichia coli -infected individuals suggests a multifaceted mechanism of virulence and antimicrobial resistance, where the cytotoxic, beta-lactamase, and adhesion-related factors may collectively contribute to the severity and persistence of urinary tract infections caused by this pathogen.
This case- control study was done at patients in hospital in Karbala province,. All patients collected were registered in UTI center in hospital from October (2023) to end of march (2024). Patients: 55 patients randomly recruited from the patients hospital in Karbala aged ranged between (20-63) years, who are diagnosed to have UTI based on clinical and laboratory findings)lymphocyte, neutrophils, eiosinophils) by the clinicians; the patients data collection were include residence area, Diabetes mellitus and gender.
One hundred fifty (150) participants were enrolled in this case control study. The first group includes 95 patients, females (50) and males (45),the second group includes 55 (29) males and (26) females as a healthy control group. All of the groups’ ages range from 20 to 63 years.All specimens were analyzed by general urine examination (GUE) to identify the presence of bacterial cell, white blood cell, and other substances. For quantitative culture and presumptive identification from each urine specimen
Well-mixed urine specimens were cultured on MacConkey agar, and EMB and chromogenic agar orientation plates, separately. The DNA of E.coli isolates were extracted and PCR technique were used to detect some virulence factor genes CNF, A/E, BLA OXA-48, BLA CTX-M , that responsible for ESBL-Producing E.coli pathogenicity.Heparin binding protein (HBP) was measurement by using Enzyme linked Immonosorbent assay for patients and control groups.The results showed the bacterial culture and the majority was 57.90% of samples culture having a bacterial growth, while 42.10% only presented with no growth. The highest percentage of bacteria isolated from patients was Escherichia coli at 70.90%, while the lowest percentage of bacteria isolated from patients was Acinetobacter baumannii and Proteus mirabilis at 3.60%. The results of the statistical analysis indicate a significant increase in HBP concentration in patients compared to the healthy control group. In the patient group, the average HBP concentration was 22.04 ± 0.96 ng/ml. In contrast, the healthy control group had an average HBP concentration of 7.78 ± 0.59 ng/ml. Bla-CTM gene was detected in 100% of UPEC isolates, conferring resistance to β-lactam antibiotics. Bla OXA-48 gene and CNF gene were detected in 7.7% of isolates, associated with carbapenem resistance and virulence, respectively. EAE gene was detected in 20.5% of isolates, linked to the ability of E. coli to adhere to and invade host cells.
The Receiver Operative Characteristic Curve (ROC) analysis yielded a cut off value of HBP for prediction of disease activity. The overall AUC, sensitivity, and specificity for HBP were as follows: 0.936, 0.091, and 0.909, respectively.The results of current study revealed that Escherichia coli remains the protein (HBP) levels were significantly higher in patients as compared to healthy control suggesting that it may be a potential biomarker for UTI. In conclusion: the significantly higher levels of HBP observed in infected patients compared to healthy controls indicate that HBP may serve as a potential biomarker for the diagnosis and monitoring of UTIs.