MOLECULAR CHARACTERIZATION AND MUTATIONAL ANALYSIS OF AMINOGLYCOSIDES, SULFONAMIDES AND QUINOLONES RESISTANT GENES OF ESCHERICHIA COLI ISOLATED FROM UTIS PATIENTS
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Keywords
E. coli, antibiogram, PCR, resistant genes, NGS, mutational analysis
Abstract
Antibiotic resistance produced by Gram-negative bacteria causing UTIs is an issue of concern for clinicians which limits the treatment options. The current study is designed to determine the molecular characterization and mutational analysis of antibiotic resistant genes of E. coli isolated from UTIs patients. A total of 93 E. coli isolates were obtained from UTIs patients at Khyber Teaching Hospital (KTH), Peshawar. The collected samples were identified phenotypically using API 20E strips. The antibiotic resistant genes of E. coli were detected by PCR and subsequently sequenced by Next Generation Sequencing (NGS) for mutational analysis. The antibiogram results revealed that most of the E. coli isolates were resistant against antibiotics; Ampicillin, cephalosporins, cotrimoxazole, aminoglycosides and ciprofloxacin while showed good results against TGC, MEM, TZP and SCF. Molecular analysis showed that 21 isolates were positive for Sul1 gene, Aac1 10, Qnrs 7 and Sul2 gene 6. The mutational analysis of resistant genes showed that Aac1 gene have two amino acid substitutions (M120T and R197I) while Qnrs, Sul1 and Sul2 have single amino acid substitution (H95R, H292L and E66A), respectively. Different mutations in antibiotic resistant genes; Aac1, Qnrs, Sul1 and Sul2 and EaeA of E. coli prove that bacteria have a drastic internal machinery modification which can lead to antibiotic resistance.
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Mohammed Haroon
Department of Medicine, Khyber Teaching Hospital, Peshawar, Pakistan.
Abdul Salam
Centre of Biotechnology and Microbiology, University of Peshawar, Pakistan.
Sadiq Azam
Centre of Biotechnology and Microbiology, University of Peshawar, Pakistan.
Noor Rehman
Centre of Biotechnology and Microbiology, University of Peshawar, Pakistan.
Department of Pathology, Khyber Teaching Hospital, Peshawar, Pakistan.
Ibrar khan
Centre of Biotechnology and Microbiology, University of Peshawar, Pakistan.
Muhammad Asghar
Department of Pathology, Khyber Teaching Hospital, Peshawar, Pakistan.
Nauman Khan
Centre of Biotechnology and Microbiology, University of Peshawar, Pakistan.
Muhammad Absar
Department of Pathology/Microbiology, King Abdulaziz Hospital, Saudi Arabia
Aakash Ahmad Khattak
Centre of Biotechnology and Microbiology, University of Peshawar, Pakistan.