Multiplex System: Identification of Vancomycin (Vana) And Methicillin (Meca) Resistance Genes In Staphylococcus Aureus

Authors

  • Dheeraj Kumar Department of Bioscience, Faculty of Life Sciences, Shaheed Zulfikar Ali Bhutto Institute of Science and Technology (SZABIST), Karachi, Pakistan
  • Waqas Ahmad Abbasi Department of Bioscience, Faculty of Life Sciences, Shaheed Zulfikar Ali Bhutto Institute of Science and Technology (SZABIST), Karachi, Pakistan.
  • Uroosa Khalil Department of Bioscience, Faculty of Life Sciences, Shaheed Zulfikar Ali Bhutto Institute of Science and Technology (SZABIST), Karachi, Pakistan.
  • Sumaiya Kazi Ziauddin Medical College, Ziauddin University, Karachi, Pakistan.
  • Hiba Saleem Medical Lab Technology, Bahria University College of Allied Health Sciences, Bahria University of Health Sciences, Karachi, Pakistan
  • Kiran Fatima Department of Bioscience, Faculty of Life Sciences, Shaheed Zulfikar Ali Bhutto Institute of Science and Technology (SZABIST), Karachi, Pakistan
  • Syed Aqib Ali Zaidi Shenzhen Key Laboratory of Anti-Aging and Regenerative Medicine, Medical School, Shenzhen University, Shenzhen, 518107, China.
  • Kashif Ali Department of Bioscience, Faculty of Life Sciences, Shaheed Zulfikar Ali Bhutto Institute of Science and Technology (SZABIST), Karachi, Pakistan.
  • Faria Fatima College of Medical Technology, Ziauddin University, Karachi Pakistan.

DOI:

https://doi.org/10.46568/bios.v4i4.146

Keywords:

Staphylococcus aureus, Resistance genes, Antibiotic Resistance, Multiplex PCR

Abstract

Introduction: Emergence of Methicillin resistant Staphylococcus aureus (MRSA) and Vancomycin resistant Staphylococcus aureus (VRSA) strain from different regions of the world poses a grave concern to human health. Antibiotic resistance in S. aureus is mainly because of the genetic factors which modify or disrupt their target site on bacteria. Methods: This study is focused to identify the vancomycin and methicillin resistance gene in antibiotic sensitive and resistant S. aureus. The pure cultures of S. aureus were isolated, subjected to morphological and biochemical characterization. Antibiotic susceptibility testing was done to check the resistance pattern. DNA isolation was followed by genotyping of the antibiotic resistance genes (VanA and MecA) and housekeeping gene (AroE) was done through multiplex PCR method. All the strains showed the colonial, microscopic and biochemical characteristics (catalase and coagulase positive) specific for S. aureus. Results: Majority of the strains were resistant to cefixime (80 %) and least resistance was observed with fusidic acid (0%), while resistance frequency of the remaining antibiotics falls between them. All the strains showed the presence of housekeeping AroE gene with frequency of VanA is 2% and MecA is 24% which coincides with the findings of antibiotic resistance testing. For VanA, there might be other resistance genes of vancomycin cassette which confer the resistance against it. Conclusion: The study will help to discriminate the vancomycin and methicilin sensitive and resistance strains of S. aureus based on their respective genetic factors and help to validate the underlying mechanism in the acquisition of antibiotic resistance.

References

Abrar S, Hussain S, Khan RA, Ul Ain N, Haider H, Riaz SJAR, et al. Prevalence of extended-spectrum-β-lactamase-producing Enterobacteriaceae: first systematic meta-analysis report from Pakistan. 2018;7(1):1-11.

Kobayashi SD, Malachowa N, DeLeo FRJTAjop. Pathogenesis of Staphylococcus aureus abscesses. 2015;185(6):1518-27.

(WHO) WHO. Antimicrobial Resistance World Health Organization (WHO); 2023 [updated 31.7.2020. Available from: https://www.who.int/news-room/fact-sheets/detail/antibiotic-resistance.

Organization WH. Global antimicrobial resistance and use surveillance system (GLASS) report: 2021. 2021.

Chinemerem Nwobodo D, Ugwu MC, Oliseloke Anie C, Al‐Ouqaili MT, Chinedu Ikem J, Victor Chigozie U, et al. Antibiotic resistance: The challenges and some emerging strategies for tackling a global menace. 2022;36(9):e24655.

Giedraitienė A, Vitkauskienė A, Naginienė R, Pavilonis AJM. Antibiotic resistance mechanisms of clinically important bacteria. 2011;47(3):19.

Munita J, Arias C. Mechanisms of antibiotic resistance. Microbiol Spectr 4: 1–37. 2016.

Van Hoek AH, Mevius D, Guerra B, Mullany P, Roberts AP, Aarts HJJFim. Acquired antibiotic resistance genes: an overview. 2011;2:203.

Gram CJFdM. The differential staining of Schizomycetes in tissue sections and in dried preparations. 1884;2(6):185-9.

O'Toole GAJJob. Classic spotlight: How the gram stain works. Am Soc Microbiol; 2016. p. 3128-.

Gagnon M, Hunting W, Esselen WJAC. New method for catalase determination. 1959;31(1):144-6.

Hensel A, Petzoldt K. Biological and biochemical analysis of bacteria and viruses. Bioaerosols Handbook: CRC Press; 2020. p. 335-60.

Sanders ERJJ. Aseptic laboratory techniques: plating methods. 2012(63):e3064.

Sperber Wz, Tatini SJAm. Interpretation of the tube coagulase test for identification of Staphylococcus aureus. 1975;29(4):502-5.

Bauer AJAJCP. Antibiotic susceptibility testing by a standardized single diffusion method. 1966;45:493-6.

Sneat P, Mair N, Sharpe M, JG HJB, London, Los Angeles, Sydney. Bergely’s Manual of Systematic Bacteriology Volume 2, Williams and Wilkins. 1986.

Masoodi KZ, Lone SM, Rasool RS. Advanced methods in molecular biology and biotechnology: a practical lab manual: Academic Press; 2020.

Enright MC, Day NP, Davies CE, Peacock SJ, Spratt BGJJocm. Multilocus sequence typing for characterization of methicillin-resistant and methicillin-susceptible clones of Staphylococcus aureus. 2000;38(3):1008-15.

Shekarabi M, Hajikhani B, Salimi Chirani A, Fazeli M, Goudarzi MJPo. Molecular characterization of vancomycin-resistant Staphylococcus aureus strains isolated from clinical samples: A three year study in Tehran, Iran. 2017;12(8):e0183607.

Hussein SAJIJVS. Study of Staphylococcus aureus isolated from the mouth of canary. 2020;34(2):301-4.

Kadhem H, Abed Ali A, Hassan OJWJEB. Isolation and identification of bacteria isolated from different parts of cell phones. 2016;4(1):29-31.

Taylor TA, Unakal CG. Staphylococcus aureus. StatPearls [Internet]: StatPearls Publishing; 2022.

van Teeseling MC, de Pedro MA, Cava FJFim. Determinants of bacterial morphology: from fundamentals to possibilities for antimicrobial targeting. 2017:1264.

Boyanova LJPm. Direct Gram staining and its various benefits in the diagnosis of bacterial infections. 2018;130(1):105-10.

Faron ML, Ledeboer NA, Buchan BWJJocm. Resistance mechanisms, epidemiology, and approaches to screening for vancomycin-resistant Enterococcus in the health care setting. 2016;54(10):2436-47.

Phukan C, Lahkar M, Ranotkar S, Saikia KKJTIJoMR. Emergence of vanA gene among vancomycin-resistant enterococci in a tertiary care hospital of North-East India. 2016;143(3):357.

Al-Amery K, Elhariri M, Elsayed A, El-Moghazy G, Elhelw R, El-Mahallawy H, et al. Vancomycin-resistant Staphylococcus aureus isolated from camel meat and slaughterhouse workers in Egypt. 2019;8:1-8.

Hussain SF, Khatoon A, Shahid SM, Ismail M, Azhar AJAJMR. Discriminative multiplex (hexaplex) PCR strategy for the detection of methicillin resistance and virulence factors in Staphylococcus aureus. 2013;7(3):196-201.

Sadiq A, Samad M, Basharat N, Ali S, Saad Z, Khan AN, et al. Methicillin-Resistant Staphylococcus aureus (MRSA) in Slaughter Houses and Meat Shops in Capital Territory of Pakistan During 2018–2019. 2020;11:577707.

Schmitz F-J, Krey A, Sadurski R, Verhoef J, Milatovic D, Fluit ACJJoac. Resistance to tetracycline and distribution of tetracycline resistance genes in European Staphylococcus aureus isolates. 2001;47(2):239-40.

Lakhundi S, Zhang KJCmr. Methicillin-resistant Staphylococcus aureus: molecular characterization, evolution, and epidemiology. 2018;31(4):e00020-18.

Mou X-B, Ali Z, Li B, Li T-T, Yi H, Dong H-M, et al. Multiple genotyping based on multiplex PCR and microarray. 2016;27(11):1661-5.

Syal K, Mo M, Yu H, Iriya R, Jing W, Guodong S, et al. Current and emerging techniques for antibiotic susceptibility tests. 2017;7(7):1795.

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Published

2023-10-10

How to Cite

Dheeraj Kumar, Waqas Ahmad Abbasi, Uroosa Khalil, Sumaiya Kazi, Hiba Saleem, Kiran Fatima, … Faria Fatima. (2023). Multiplex System: Identification of Vancomycin (Vana) And Methicillin (Meca) Resistance Genes In Staphylococcus Aureus. BioSight, 4(4), 9–16. https://doi.org/10.46568/bios.v4i4.146

Issue

Vol. 4 No. 4 (2023): October

Section

Original Article

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