MOLECULAR CHARACTERIZATION AND MUTATIONAL ANALYSIS OF AMINOGLYCOSIDES, SULFONAMIDES AND QUINOLONES RESISTANT GENES OF ESCHERICHIA COLI ISOLATED FROM UTIS PATIENTS

Main Article Content

Mohammed Haroon
Abdul Salam
Sadiq Azam
Noor Rehman
Ibrar khan
Muhammad Asghar
Nauman Khan
Muhammad Absar
Aakash Ahmad Khattak

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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References

1. Adib N, Ghanbarpour R, Solatzadeh H, Alizade H (2014). Antibiotic resistance profile and virulence genes of uropathogenic E. coli isolates in relation to phylogeny. Trop Biomed., 31(1): 17-25.
2. Al-Badr A, Al-Shaikh G (2013). Recurrent urinary tract infections management in women: a review. Sultan Qaboos University Medical Journal., 13(3): 359.
3. Asokan GV, Vanitha A (2017). Disaster response under One Health in the aftermath of Nepal earthquake, 2015. Journal of epidemiology and global health., 7(1): 91-96.
4. Bonelli RR, Moreira BM, Picão RC (2014). Antimicrobial resistance among Enterobacteriaceae in South America: history, current dissemination status and associated socioeconomic factors. Drug Resistance Updates., 17(1-2): 24-36.
5. Brauner A, Katouli M, Östenson C (1995). P-fimbriation and haemolysin production are the most important virulence factors in diabetic patients with Escherichia coli bacteraemia: a multivariate statistical analysis of seven bacterial virulence factors. Journal of Infection., 31(1): 27-31.
6. Can F, Azap OK, Seref C, Ispir P, Arslan H, Ergonul O (2015). Emerging Escherichia coli O25b/ST131 clone predicts treatment failure in urinary tract infections. Clinical Infectious Diseases., 60(4): 523-527.
7. Cattoir V, Poirel L, Nordmann P (2007). In-vitro mutagenesis of qnrA and qnrS genes and quinolone resistance in Escherichia coli. Clinical Microbiology and Infection., 13(9): 940-943.
8. Foxman B (2014). Urinary tract infection syndromes: occurrence, recurrence, bacteriology, risk factors, and disease burden. Infectious Disease Clinics., 28(1): 1-13.
9. Gonçalves LF, de Oliveira Martins-Júnior P, de Melo ABF, da Silva RCRM, de Paulo Martins V, Pitondo-Silva A, et al. (2016). Multidrug resistance dissemination by extended-spectrum β-lactamase-producing Escherichia coli causing community-acquired urinary tract infection in the Central-Western Region, Brazil. Journal of Global Antimicrobial Resistance., 6: 1-4.
10. Gupta K, Hooton TM, Stamm WE (2001). Increasing antimicrobial resistance and the management of uncomplicated community-acquired urinary tract infections. Annals of internal medicine., 135(1): 41-50.
11. Gurevich E, Tchernin D, Schreyber R, Muller R, Leibovitz E (2016). Follow-up after infants younger than 2 months of age with urinary tract infection in Southern Israel: epidemiologic, microbiologic and disease recurrence characteristics. Brazilian Journal of Infectious Diseases., 20: 19-25.
12. Hammerum AM, Sandvang D, Andersen SR, Seyfarth AM, Porsbo LJ, Frimodt-Møller N, et al. (2006). Detection of sul1, sul2 and sul3 in sulphonamide resistant Escherichia coli isolates obtained from healthy humans, pork and pigs in Denmark. International journal of food microbiology., 106(2): 235-237.
13. Harding G, Ronald A (1994). The management of urinary infections; what have we learned in the past decade? International journal of antimicrobial agents., 4(2): 83-88.
14. Holmberg SD, Solomon SL, Blake PA (1987). Health and economic impacts of antimicrobial resistance. Reviews of infectious diseases., 9(6): 1065-1078.
15. Johnson JR, Stell AL (2000). Extended virulence genotypes of Escherichia coli strains from patients with urosepsis in relation to phylogeny and host compromise. The Journal of infectious diseases., 181(1): 261-272.
16. Karishetti MS, Shaik HB (2019). Clinicomicrobial assessment of urinary tract infections in a tertiary care hospital. Indian Journal of Health Sciences and Biomedical Research., 12(1): 69.
17. Kärkkäinen U, Ikäheimo R, Katila M, Sivonen A, Siitonen A (2000). Low virulence of Escherichia coli strains causing urinary tract infection in renal disease patients. European Journal of Clinical Microbiology and Infectious Diseases., 19(4): 254-259.
18. Karlowsky JA, Kelly LJ, Thornsberry C, Jones ME, Sahm DF (2002). Trends in antimicrobial resistance among urinary tract infection isolates of Escherichia coli from female outpatients in the United States. Antimicrobial Agents and Chemotherapy., 46(8): 2540-2545.
19. Kurazono H, Nakano M, Yamamoto S, Ogawa O, Yuri K, Nakata K, et al. (2003). Distribution of the usp gene in uropathogenic Escherichia coli isolated from companion animals and correlation with serotypes and size‐variations of the pathogenicity island. Microbiology and immunology., 47(10): 797-802.
20. Leimbach A, Hacker J, Dobrindt U (2013). E. coli as an all-rounder: the thin line between commensalism and pathogenicity. Between pathogenicity and commensalism: 3-32.
21. Majeed A, Moser K (1999). Age-and sex-specific antibiotic prescribing patterns in general practice in England and Wales in 1996. British Journal of General Practice., 49(446): 735-736.
22. O’Brien VP, Hannan TJ, Schaeffer AJ, Hultgren SJ (2015). Are you experienced? Understanding bladder innate immunity in the context of recurrent urinary tract infection. Current opinion in infectious diseases., 28(1): 97.
23. Odoki M, Almustapha Aliero A, Tibyangye J, Nyabayo Maniga J, Wampande E, Drago Kato C, et al. (2019). Prevalence of bacterial urinary tract infections and associated factors among patients attending hospitals in Bushenyi district, Uganda. International journal of microbiology., 2019.
24. Planta MB (2007). The role of poverty in antimicrobial resistance. The Journal of the American Board of Family Medicine., 20(6): 533-539.
25. Prats G, Navarro F, Mirelis B, Dalmau D, Margall N, Coll P, et al. (2000). Escherichia coli serotype O15: K52: H1 as a uropathogenic clone. Journal of Clinical Microbiology., 38(1): 201-209.
26. Reller LB, Weinstein M, Jorgensen JH, Ferraro MJ (2009). Antimicrobial susceptibility testing: a review of general principles and contemporary practices. Clinical Infectious Diseases., 49(11): 1749-1755.
27. Röderova M, Halova D, Papousek I, Dolejska M, Masarikova M, Hanulik V, et al. (2017). Characteristics of quinolone resistance in Escherichia coli isolates from humans, animals, and the environment in the Czech Republic. Frontiers in microbiology., 7: 2147.
28. Tullus K, Hörlin K, Svenson SB, Källenius G (1984). Epidemic outbreaks of acute pyelonephritis caused by nosocomial spread of P fimbriated Escherichia coli in children. Journal of Infectious Diseases., 150(5): 728-736.

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