Duplex PCR detection of blashv and blatem genes in uropathogenic escherichia coli (UPEC) isolates from patient

Lee, Yin Ying (2022) Duplex PCR detection of blashv and blatem genes in uropathogenic escherichia coli (UPEC) isolates from patient. Final Year Project, UTAR.

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Abstract

The emergence of multidrug-resistant and extended spectrum beta-lactamases-producing uropathogenic Escherichia coli (ESBLs-producing UPEC), which is the leading aetiological agent of urinary tract infections (UTIs), causing a significant reduction in the efficacy of antimicrobial treatment and increase in mortality. This study aimed to investigate the prevalence of blaSHV and blaTEM genes, and to determine the antibiotic resistance profile among the UPEC isolates as well as the association between the phenotypic and genotypic data. UPEC strains (n=60) were isolated from patients’ urine samples. The isolates had undergone bacterial strain identification using MALDI-TOF by our collaborator in Hospital Raja Permaisuri Bainun in Ipoh. Kirby-Bauer disk diffusion was performed with nine antimicrobial agents that come from five different classes to determine the antibiotic resistance profile. The fast boil method was carried out for DNA extraction from the UPEC isolates. To screen the blaSHV and blaTEM genes, duplex PCR was conducted. The antibiotic susceptibility result showed that 71.67% of the isolates were resistant to ampicillin, 51.67% to tetracycline, 50.00% to nalidixic acid, 33.33% to SXT, 31.67% to ciprofloxacin, 26.67% to levofloxacin, 16.67% to chloramphenicol, 1.67% to minocycline, while all the isolates were susceptible to imipenem. The molecular result revealed that blaTEM gene was detected in most of the UPEC isolates (60.00%, n=36) while none of the isolates was detected with the presence of blaSHV gene, suggesting that there are variants of blaSHV gene that may not be able to detect in this study. The positive associations between blaTEM gene and four of the tested antibiotics: ampicillin (p=0.014), ciprofloxacin (p=0.041), levofloxacin (p=0.043) and tetracycline (p=0.001), were found to be significant. Distribution of blaTEM was revealed to be predominant in female patients and in patients who aged 60 years and above. However, negative association was observed between the blaTEM genes and gender as well as age.

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