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Prevalence and Evaluation of Toxin Genes among Uropathogenic Escherichia coli Clinical Isolates by Duplex PCR

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Abstract:

Background: One of the most common infections in human is urinary tract infection (UTI) and Uropathogenic Escherichia coli is one of its major causative agents. UTI is extremely common among young women. Children under age 5 are also highly at risk. Considering the prevalence of this disease, it is necessary to design an appropriate diagnostic method for its effective diagnosis. The aim of present study was to identify the prevalence of two virulence genes (sat and vat) among Uropathogenic E. coli isolates. Methods: Urine samples were taken from 350 patients with urinary tract infection. The samples were cultured on EMB agar and Blood agar. The suspected E. coli colonies were isolated and confirmed by biochemical tests. The genomic DNA was extracted from 297 isolated E. coli and target genes were amplified by PCR. The amplicons were sequenced and analyzed with ClustalW software. Moreover, data analysis was performed by using SPSS software. Subsequently, Duplex PCR was optimized for simultaneous detection of two genes. Results: The prevalence of sat and vat genes were 75 (n: 225) and 36 (n: 106) percent, respectively. In addition, less than 4% (n: 11) of clinical isolates comprised two genes. Conclusion: According to the conducted research, molecular identification of Uropathogenic E .coli strains according to detection of sat gene is potentially an appropriate method and could be noted for diagnosis.

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