Antibiogram analysis and molecular characterization of multi-drug resistant genes (tetk and gyra) of staphylococcus aureus isolated from diabetic foot ulcers patients in peshawar

Khan Kamran Khan; Aman Ullah; Ihtisham UL Haq; Omer Khiam; Zaryab Khan; Musa Khan

doi:10.22376/

Authors

Kamran Khan Department of Health and Biological Sciences, Abasyn University, Peshawar, Pakistan
Aman Ullah Institute of Biotechnology and Genetics Engineering, University of Agriculture Peshawar, Pakistan
Ihtisham UL Haq Department of Rehabilitation & Allied Health Sciences, Riphah International University, Pakistan
Omer Khiam Institute of Biotechnology and Genetics Engineering, University of Agriculture Peshawar, Pakistan
Zaryab Khan Department of Nursing, Zalan Collage mingora swat, Pakistan
Musa Khan Gulrang Institute of Nursing Thana Malakand, Pakistan

Abstract

DFUs are one of the frequent complications of diabetic patients frequently resulting in the emergence of bacterial infections that make treatment difficult. The aim of this research was aimed at determining the antibiogram and molecular characterization of multidrug-resistant (MDR) genes, namely tetK and gyrA, in Staphylococcus aureus that was isolated on DFU patients in Peshawar, Pakistan. Sixty wound samples were taken out of DFU patients of Lady Reading Hospital, where more than 70% of respondents were males. Bacterial identification was done using biochemical tests (catalase and coagulase) and the samples were grown on the Mannitol Salt Agar (MSA). The Kirby-Bauer disc diffusion was utilized to identify the antibiotic susceptibility and PCR was done to identify the tetK and gyrA resistance genes. These findings indicated that rates of resistance to popular antibiotics were high with 100 percent resistance to ampicillin (100%), penicillin G (93%), and erythromycin (83%), with intermediate rates of resistance to tetracycline and ciprofloxacin. Further, the proportion of methicillin-resistant S. aureus (MRSA) and oxacillin-resistant of the isolates was 48 and 83 percent respectively. Molecular testing confirmed the existence of gyrA gene in 15 samples and tetK gene in 14 samples in which the two genes were important in antibiotic resistance. To sum up, the present paper highlights the increasing problem of antibiotic resistance to DFU infection. It highlights that persistent molecular monitoring and individualized treatment approaches should be used to provide effective treatment of infections among diabetic individuals.

Keywords:

S. aureus, tetK, gyrA, resistant genes, Resistant antibiotics

DOI

https://doi.org/10.22376/

References

1. Rawshani A, Rawshani A, Franzén S, Eliasson B, Svensson AM, Miftaraj M, et al. Mortality and cardiovascular disease in type 1 and type 2 diabetes. N Engl J Med. 2017 Apr 13;376(15):1407–18.

2. Alam U, Asghar O, Azmi S, Malik RA. General aspects of diabetes mellitus. Handb Clin Neurol. 2014;126:211–22.

3. Lima MJ, Domingues MR, Crowley VM, Sousa MI, Monteiro PA, Leandro CI, et al. Pathogenesis of Type 2 Diabetes Mellitus: The Multifaceted Roles of Insulin Resistance, β-Cell Dysfunction, and Inflammation. Cells. 2022;11(2):312.

4. Saeed A, Ahsan F, Nawaz M, Iqbal K, Rehman KU, Ijaz T. Incidence of vancomycin resistant phenotype of the methicillin resistant Staphylococcus aureus isolated from a tertiary care hospital in Lahore. Antibiotics. 2019 Dec 18;9(1):3.

5. Hasan SU, Siddiqui MR. Nationwide prevalence of type 2 diabetes mellitus and pre-diabetes in Pakistan: A systematic review and meta-analysis. Diabetes Res Clin Pract. 2024 Oct 1;216:111815.

6. Shrivastava U, Misra A, Mohan V, Unnikrishnan R, Bachani D. Obesity, diabetes and cardiovascular diseases in India: public health challenges. Curr Diabetes Rev. 2017 Feb 1;13(1):65–80.

7. Mondal UK, Ahmed KY, Thapa S, Kalinna B, Pak SC, Anyasodor AE, et al. A systematic review of the major risk factors for type two diabetes among Aboriginal Australians. BMC Public Health. 2024 Nov 21;24(1):3174.

8. Pliszka M, Szablewski L. Associations between diabetes mellitus and selected cancers. Int J Mol Sci. 2024 Jul 8;25(13):7476.

9. Hui D, Judd AC, Moneme C, Passerini H, Silpe S, Podboy A, et al. Synergistic Integration of Multimodal Metabolic and Bariatric Interventions Transforming Transplant Care. J Clin Med. 2025 Aug 11;14(16):5669.

10. de Oliveira LE. Staphylococcus aureus and Septic Arthritis of the Knee: Diagnostic and Antimicrobial Susceptibility [Master’s thesis]. Porto (Portugal): Universidade do Porto; [no date].

11. Khan D, Zeb M, Khattak SK, Shah AA, Abdullah M, Bilal M. Molecular characterization and antibiotic susceptibility pattern of bacterial strains isolated from diabetic foot ulcers. Endocr Metab Sci. 2023 Sep 30;12:100136.

12. Narendrakumar L, Chakraborty M, Kumari S, Paul D, Das B. β-Lactam potentiators to re-sensitize resistant pathogens: Discovery, development, clinical use and the way forward. Front Microbiol. 2023 Mar 10;13:1092556.

13. Aboelnaga N, Elsayed SW, Abdelsalam NA, Salem S, Saif NA, Elsayed M, et al. Deciphering the dynamics of methicillin-resistant Staphylococcus aureus biofilm formation: from molecular signaling to nanotherapeutic advances. Cell Commun Signal. 2024 Mar 22;22(1):188.

14. Mattsson K, Pihlsgård M, Enhörning S, Timpka S. Incident cardiovascular disease in women with type 1 or type 2 diabetes following a hypertensive disorder of pregnancy. Hypertension. 2024 Apr;81(4):897–905.

15. Huband MD, Thompson JD, Gurung ND, Liu Q, Li L, Zhang J, et al. Activity of the novel aminomethylcycline KBP-7072 and comparators against 1,057 geographically diverse recent clinical isolates from the SENTRY surveillance program, 2019. Antimicrob Agents Chemother. 2022 Jan 18;66(1):e01397-21.

16. Mansoor Z, Modaweb A. Predicting amputation in patients with diabetic foot ulcers: a systematic review. Cureus. 2022 Jul 25;14(7):e27363.

17. Saleem M, Ahmad I, Salem AM, Almarshedy SM, Moursi SA, Syed Khaja AS, et al. Molecular and genetic analysis of methicillin-resistant Staphylococcus aureus (MRSA) in a tertiary care hospital in Saudi Arabia. Naunyn Schmiedebergs Arch Pharmacol. 2025 Jan 8:1–1.

18. Walia S. Prevalence of diabetic retinopathy among self-reported newly diagnosed diabetics. Igmin Res. 2024 May 16;2(5):370–3.

19. Huang X, Li G. Antimicrobial peptides and cell-penetrating peptides: non-antibiotic membrane-targeting strategies against bacterial infections. Infect Drug Resist. 2023 Dec 31:1203–19.

20. Chowdhury ME, Zughaier SM, Hasan A, Alfkey R, editors. Diabetic Foot Ulcers - Pathogenesis, Innovative Treatments and AI Applications. Norderstedt: BoD – Books on Demand; 2024 May 22.

21. Porte L, Soto A, Andrighetti D, Dabanch J, Braun S, Saldivia A, et al. Catheter-associated bloodstream infection caused by Leifsonia aquatica in a haemodialysis patient: a case report. J Med Microbiol. 2012 Jun;61(6):868–73.

22. Suh K, Toye B, Jessamine P, Chan F, Ramotar K. Epidemiology of methicillin-resistant Staphylococcus aureus in three Canadian tertiary-care centers. Infect Control Hosp Epidemiol. 1998 Jul;19(6):395–400.

23. Tacconelli E, Carrara E, Savoldi A, Harbarth S, Mendelson M, Monnet DL, et al. Discovery, research, and development of new antibiotics: the WHO priority list of antibiotic-resistant bacteria and tuberculosis. Lancet Infect Dis. 2018 Mar 1;18(3):318–27.

24. Tanaka M, Wang T, Onodera Y, Uchida Y, Sato K. Mechanism of quinolone resistance in Staphylococcus aureus. J Infect Chemother. 2000 Sep;6(3):131–9.

25. Thaker M, Spanogiannopoulos P, Wright GD. The tetracycline resistome. Cell Mol Life Sci. 2010 Feb;67(3):419–31.

26. Song M. Cancer overtakes vascular disease as leading cause of excess death associated with diabetes. Lancet Diabetes Endocrinol. 2021 Mar 1;9(3):131–3.

27. Tong SY, Davis JS, Eichenberger E, Holland TL, Fowler VG Jr. Staphylococcus aureus infections: epidemiology, pathophysiology, clinical manifestations, and management. Clin Microbiol Rev. 2015 Jul;28(3):603–61.

28. Trzcinski K, Cooper BS, Hryniewicz W, Dowson CG. Expression of resistance to tetracyclines in strains of methicillin-resistant Staphylococcus aureus. J Antimicrob Chemother. 2000 Jun 1;45(6):763–70.

29. Turner NA, Sharma-Kuinkel BK, Maskarinec SA, Eichenberger EM, Shah PP, Carugati M, et al. Methicillin-resistant Staphylococcus aureus: an overview of basic and clinical research. Nat Rev Microbiol. 2019 Apr;17(4):203–18.

30. Ullah N, Dar HA, Naz K, Andleeb S, Rahman A, Saeed MT, et al. Genomic investigation of methicillin-resistant Staphylococcus aureus ST113 strains isolated from tertiary care hospitals in Pakistan. Antibiotics. 2021 Sep 17;10(9):1121.