Identification of Gene Mutation Patterns Obtained from Resistance to Mupirocin in Methicillin-Resistant Staphylococcus Aureus Clinical Strains, Using High-Resolution Melting (HRM) Method

Document Type : Original Article (s)

Authors

1 Department of Microbiology, School of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran

2 Department of Microbiology, School of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran

3 Associate Professor, Brucellosis Research Center AND Department of Microbiology, School of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran

Abstract

Background: Effective antibiotics on the translation pathway, such as mupirocin, may cause gene mutations in methicillin-resistant Staphylococcus aureus in long-term. Using high-sensitivity methods plays an important role in identifying these bacteria. Our goal was to identify these mutations using high-resolution melting (HRM) curve of DNA analysis.Methods: Resistance to mupirocin was identified using disc microdilution plate method in according to the Clinical and Laboratory Standards Institute (CLSI) guideline. mupA gene amplification in the isolates of methicillin-resistant Staphylococcus aureus was done using polymerase chain reaction (PCR). Then, analysis was performed using StepOne Software and HRM software. Sequencing was used as gold-standard method for confirming of the results.Findings: Out of 162 Staphylococcus aureus isolates, 83 (51.32%) were methicillin resistant. Among methicillin-resistant Staphylococcus aureus isolates, 47 (52.80 %) showed high level resistance to mupirocin carrying mupA. The most and lowest resistance was observed for penicillin (79.62%) and ceftazidime (6.17%), respectively. Moreover, all of multi-drug resistant (MDR) isolates were mupirucin resistant, too. Among mupirocin-resistant Staphylococcus aureus, wound samples were the most prevalent. Besides, isolates obtained from wound and blood demonstrated the highest mutation in A and G bases. Meaningful association was observed between the type of clinical sample and mutation rate, and resistance to mupirocin and methicillin (P < 0.05).Conclusion: Mupirocin-derived gene mutations provide multi-drug resistance in methicillin-resistant Staphylococcus aureus.

Keywords

References

  1. Walter J, Noll I, Feig M, Weiss B, Claus H, Werner G, et al. Decline in the proportion of methicillin resistance among Staphylococcus aureus isolates from non-invasive samples and in outpatient settings, and changes in the co-resistance profiles: An analysis of data collected within the Antimicrobial Resistance Surveillance Network, Germany 2010 to 2015. BMC Infect Dis 2017; 17(1): 169.
  2. Sweeney NL, Lipker L, Hanson AM, Bohl CJ, Engel KE, Kalous KS, et al. Docking into Mycobacterium tuberculosis thioredoxin reductase protein yields pyrazolone lead molecules for methicillin-resistant Staphylococcus aureus. Antibiotics (Basel) 2017; 6(1).
  3. Tahmasebi H, Zeyni B, Dehbashi S, Motamedi H, Vafaeefar M, Keramat F, et al. The study of blaZ and mecA gene expression in methicillin-resistant Staphylococcus aureus strains and the relationship between the gene expression patterns. J Isfahan Med Sch 2017; 35(443): 1062-7. [In Persian].
  4. Sussmuth RD, Mainz A. nonribosomal peptide synthesis-principles and prospects. Angew Chem Int Ed Engl 2017; 56(14): 3770-821.
  5. Caffrey AR, Quilliam BJ, LaPlante KL. Risk factors associated with mupirocin resistance in meticillin-resistant Staphylococcus aureus. J Hosp Infect 2010; 76(3): 206-10.
  6. Hesami S, Hosseini SD, Amouzandeh-Nobaveh A, Eskandari S, Ghaznavi-Rad E. Phenotypic and genotypic determination of mupirocin resistance among methicillin susceptibility and resistance in staphylococci isolated from nosocomial infections. J Mazandaran Univ Med Sci 2014; 23(1): 30-9. [In Persian].
  7. Stefanaki C, Ieronymaki A, Matoula T, Caroni C, Polythodoraki E, Chryssou SE, et al. Six-year retrospective review of hospital data on antimicrobial resistance profile of Staphylococcus aureus isolated from skin infections from a single institution in GREECE. Antibiotics (Basel) 2017; 6(4).
  8. International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements (IWG-SCC). Classification of staphylococcal cassette chromosome mec (SCCmec): Guidelines for reporting novel SCCmec elements. Antimicrob Agents Chemother 2009; 53(12): 4961-7.
  9. Fritz SA, Hogan PG, Camins BC, Ainsworth AJ, Patrick C, Martin MS, et al. Mupirocin and chlorhexidine resistance in Staphylococcus aureus in patients with community-onset skin and soft tissue infections. Antimicrob Agents Chemother 2013; 57(1): 559-68.
  10. Rajkumari N, Mathur P, Bhardwaj N, Gupta G, Dahiya R, Behera B, et al. Resistance pattern of mupirocin in methicillin-resistant Staphylococcus aureus in trauma patients and comparison between disc diffusion and E-test for better detection of resistance in low resource countries. J Lab Physicians 2014; 6(2): 91-5.
  11. Krishnan PU, Miles K, Shetty N. Detection of methicillin and mupirocin resistance in Staphylococcus aureus isolates using conventional and molecular methods: A descriptive study from a burns unit with high prevalence of MRSA. J Clin Pathol 2002; 55(10): 745-8.
  12. Hosseini SJ, Nazemi A, Hashemi M, Miri Nargesi M, Sharifi SA. Application of high resolution melting technique for detection of germ line single nucleotide polymorphisms in STK11 gene among patients with various gastrointestinal cancers. Med Sci J Islamic Azad Univ Tehran Med Branch 2012; 21(4): 233-7. [In Persian].
  13. Haryono SJ, Datasena IGB, Hariadi A, Mulyarahardj R. High resolution melting (HRM) analysis for genetic changes in BRCA1/2 gene. Supplement J Med Sci 2016; 48(4): 26.
  14. Chernukha IM, Minaev MY, Kurbakov KA, Bataeva DS. Detection and identification of S. carnosus in starter cultures using real time PCR and subsequent HRM analysis of amplification products. Procedia Food Science 2015; 5: 38-41.
  15. Clinical and Laboratory Standards Institute. Performance Standards for Antimicrobial Susceptibility Testing. CLSI supplement M100. 27th ed. Wayne, PA: CLSI; 2017.
  16. Arabestani MR, Tahmasebi H, Zeyni B. Diagnostic value of melting curve analysis based on multiplex-real time PCR in identification of enterococci species. J Mazandaran Univ Med Sci. 2017; 26(145): 234-47. [In Persian].
  17. Driscoll DG, Young CL, Ochsner UA. Transient loss of high-level mupirocin resistance in Staphylococcus aureus due to MupA polymorphism. Antimicrob Agents Chemother 2007; 51(6): 2247-8.
  18. Simor AE, Stuart TL, Louie L, Watt C, Ofner-Agostini M, Gravel D, et al. Mupirocin-resistant, methicillin-resistant Staphylococcus aureus strains in Canadian hospitals. Antimicrob Agents Chemother 2007; 51(11): 3880-6.
  19. Palepou MF, Johnson AP, Cookson BD, Beattie H, Charlett A, Woodford N. Evaluation of disc diffusion and Etest for determining the susceptibility of Staphylococcus aureus to mupirocin. J Antimicrob Chemother 1998; 42(5): 577-83.
  20. Munita JM, Arias CA. Mechanisms of antibiotic resistance. Microbiol Spectr 2016; 4(2).
  21. Hershberg R, Petrov DA. Evidence that mutation is universally biased towards AT in bacteria. PLoS Genet 2010; 6(9): e1001115.
  22. Bratchikov M, Mauricas M. Development of a multiple-run high-resolution melting assay for Salmonella spp. genotyping HRM application for Salmonella spp. subtyping. Diagn Microbiol Infect Dis 2011; 71(3): 192-200.
  23. Carbonell P, Turpin MC, Torres-Moreno D, Molina-Martinez I, Garcia-Solano J, Perez-Guillermo M, et al. Comparison of allelic discrimination by dHPLC, HRM, and TaqMan in the detection of BRAF mutation V600E. J Mol Diagn 2011; 13(5): 467-73.
Journal of Isfahan Medical School
Volume 36, Issue 476 - Serial Number 476
March and April 2018
Pages 403-410

Files

History

  • Receive Date: 23 April 2018
  • Revise Date: 01 November 2024
  • Accept Date: 06 April 2022

Share

How to cite

Statistics