Molecular characterization of blaTEM and blaCTX-M ESBLs genes producing Escherichia coli isolates from urinary tract infections (UTIs) in Al-Basrah province, Iraq

Abstract

Urinary tract infections (UTIs) are primarily caused by Gram-negative Enterobacteriaceae bacteria, accounting for 80–90% of uropathogenic Escherichia coli infections. Multidrug-resistant (MDR) bacteria, such as extended-spectrum β-lactamases (ESBLS), pose a significant threat to global healthcare systems. The current study relied on 200 urine samples collected from patients suffering urinary tract infections (UTIs) in Al Sadr Teaching Hospital in Al-Basrah Province,Iraq from the 5th January to 22nd February. A result of cultivated 200 samples collected from urinary tract infection patients showed 71 (35.5%) positive samples on MacConkey agar, eosin methylene blue (EMB), and HiChromeTM E.coli agar, and bacterial growth was distributed to 47 (66.2%) Escherichia coli and 24 (33.8%) of Gram-negative species. The diagnostic gene 16S rDNA by PCR method showed that all 47 E. coli isolates had a molecular weight of around 585 bp at 100%. Furthermore, positive results were shown in 44 (93.6%) of the E.coli isolates in the current study that produced ESBLs by using the double-disc approximation method (DAM). While 3 (6.4%) isolates revealed negative results for ESBLs, whereas the13(27.7%) of the E.coli isolates gave positive results for ESBL. While 34(72.3%) E.coli isolates showed negative results for produced ESBLs by using the double-disc synergy test (DDST), there were significant differences (P<0.01) between positive and negative isolates in both methods used to detect ESBL-producing isolates. Extended-spectrum β-lactamses (ESBLs) genes were detected by using PCR to amplify the blaTEM and blaCTX-M genes in E.coli isolates. The amplified genes' bands were characterised approximately at (800 bp) for blaTEM and (754 bp) for blaCTX-M   , the products were compared to the standard molecular DNA ladder at (2000 bp). According to the results, 47(100%) E.coli isolates gave positive results for the blaTEM gene. While blaCTX-M gene was shown, only 23(48.9%) E.coli isolates had positive results for the blaCTX-M gene.