Genotyping of Methicillin Resistant Staphylococcus aureus Clinical Isolates in Sulaimani City using Pulsed -Field Gel Electrophoresis

Main Article Content

Dunya Emad Naji
Sehand Kamaluldeen Arif

Abstract

Methicillin-resistant Staphylococcus aureus (MRSA) strains cause serious nosocomial infections. The objective of this study was to detect and find the epidemiological relatedness among MRSA isolates which was collected in Emergency burn-and-plastic surgery hospital in Sulaimani city using Pulsed-filed gel electrophoresis (PFGE). Routine methods were used for detection of Staphylococcus aureus, further molecular confirmation was done by PCR assays targeting 16S rRNA gene. The cefoxitin-disc diffusion assay was used for detection MRSA, which is subsequently confirmed by the presence of the mecA gene. PFGE was chosen to observe the genetic relatedness of 10 MRSA strains using SmaI restriction endonuclease. All the isolates (100%, 50/50) carried 16S rRNA gene specific for S.aureus and (42%, 21/50) exhibited resistance to cefoxitin and harboured the mecA gene, thereby being classified as MRSA. Among 10 MRSA isolates, the PFGE pattern were grouped into four clusters. Three isolates belonging to cluster (C3) were indistinguishable, 4 isolates belonging to cluster (C4) were closely related, 2 isolates of the second cluster (C2) were possibly related and the first cluster (C1) with 1 MRSA isolate was unrelated. This finding shows 30% of the samples had indistinguishable fingerprints with no changes in banding profiles, while 40% of the samples had closely related patterns. PFGE is a good discriminatory tool for typing MRSA strains.

Article Details

How to Cite
Naji, D. E., & Arif, S. K. (2024). Genotyping of Methicillin Resistant Staphylococcus aureus Clinical Isolates in Sulaimani City using Pulsed -Field Gel Electrophoresis. Tikrit Journal of Pure Science, 29(1), 1–15. https://doi.org/10.25130/tjps.v29i1.1397
Section
Articles

References

[1]. Algammal, A. M., Hetta, H. F., Elkelish, A., Alkhalifah, D. H., Hozzein, W. N., Batiha, G. E.-S., El Nahhas, N., & Mabrok, M. A. (2020). Methicillin-Resistant Staphylococcus aureus (MRSA): one health perspective approach to the bacterium epidemiology, virulence factors, antibiotic-resistance, and zoonotic impact. Infection and Drug Resistance, 13, 3255–3265.

[2]. Chadi, Z. D., Dib, L., Zeroual, F., & Benakhla, A. (2022). Usefulness of molecular typing methods for epidemiological and evolutionary studies of Staphylococcus aureus isolated from bovine intramammary infections. Saudi Journal of Biological Sciences, 29(8), 103338.

[3]. Moghadam, S. O., Pourmand, M. R., Douraghi, M., Sabzi, S., & Ghaffari, P. (2017). Utilization of PFGE as a powerful discriminative tool for the investigation of genetic diversity among MRSA strains. Iranian journal of public health, 46(3), 351.

[4]. Miao, J., Chen, L., Wang, J., Wang, W., Chen, D., Li, L., ... & Xu, Z. (2017). Current methodologies on genotyping for nosocomial pathogen methicillin-resistant Staphylococcus aureus (MRSA). Microbial Pathogenesis, 107, 17-28.

[5]. Rebic, V., Budimir, A., Aljicevic, M., Bektas, S., Vranic, S. M., & Rebic, D. (2016). Typing of methicillin resistant Staphylococcus aureus using DNA fingerprints by pulsed-field gel electrophoresis. Acta Informatica Medica, 24(4), 248.

[6]. Szabó, J. (2014). Molecular methods in epidemiology of Methicillin resistant Staphylococcus aureus (MRSA): advantages, disadvantages of different techniques. Journal of Medical Microbiology & Diagnosis, 3(3), 1.

[7]. PulseNet (OPN) - Centers for Disease Control and Prevention. Oxacillin¬ resistant Staphylococcus aureus on PulseNet (OPN): Laboratory Protocol for Molecular Typing of S.aureus by Pulsed¬field Gel Electrophoresis (PFGE). https://www.cdc.gov/mrsa/pdf/ar_mras_PFGE_s_aureus.pdf

[8]. Chigbu, C. O., & Ezeronye, O. U. (2003). Antibiotic resistant Staphylococcus aureus in Abia state of Nigeria. African Journal of biotechnology, 2(10), 374-378

[9]. Niyas, F. M., & Gopinath, P. (2016). Comparative study on detection of MRSA using oxacillin agar screening method, cefoxitin disc diffusion method and mecAgene by PCR among clinical isolates of Staphylococcus aureus. Research Journal of Pharmacy and Technology, 9(9), 1317-1320.

[10]. Ciftci, A., Findik, A., Onuk, E. E., & Savasan, S. (2009). Detection of methicillin resistance and slime factor production of Staphylococcus aureus in bovine mastitis. Brazilian Journal of Microbiology, 40, 254-261.

[11]. Findik, A., Akan, N., Onuk, E. E., Çakiroğlu, D., & Ciftci, A. (2009). Methicillin resistance profile and molecular typing of Staphylococcus aureus strains isolated from noses of the healthy dogs. Kafkas Univ Vet Fak Derg, 15, 925-930.

[12]. Lee, P. Y., Costumbrado, J., Hsu, C. Y., & Kim, Y. H. (2012). Agarose gel electrophoresis for the separation of DNA fragments. JoVE (Journal of Visualized Experiments), (62), e3923.

[13]. Murchan, S., Kaufmann, M. E., Deplano, A., de Ryck, R., Struelens, M., Zinn, C. E., ... & Cookson, B. (2003). Harmonization of pulsed-field gel electrophoresis protocols for epidemiological typing of strains of methicillin-resistant Staphylococcus aureus: a single approach developed by consensus in 10 European laboratories and its application for tracing the spread of related strains. Journal of clinical microbiology, 41(4), 1574-1585.

[14]. Tenover, F. C., Arbeit, R. D., Goering, R. V., Mickelsen, P. A., Murray, B. E., Persing, D. H., & Swaminathan, B. (1995). Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. Journal of clinical microbiology, 33(9), 2233-2239.

[15]. Aljanaby, A. A. J., & Aljanaby, I. A. J. (2018). Prevalence of aerobic pathogenic bacteria isolated from patients with burn infection and their antimicrobial susceptibility patterns in Al-Najaf City, Iraq-a three-year cross-sectional study. F1000Research, 7(1157), 1157.

[16]. Zahlane, K., Ouafi, A. T., & Barakate, M. (2020). The clinical and epidemiological risk factors of infections due to multi-drug resistant bacteria in an adult intensive care unit of University Hospital Center in Marrakesh-Morocco. Journal of Infection and Public Health, 13(4), 637-643.

[17]. Hussein, N., Salih, R. S., & Rasheed, N. A. (2019). Prevalence of methicillin-resistant Staphylococcus aureus in hospitals and community in Duhok, Kurdistan region of Iraq. International Journal of Infection, 6(2).1-4. https://doi.org/10.5812/iji.89636

[18]. Hasan, S. A., Saleh, I., & Ali, H. (2021). Bacteriological and Molecular Detection of Staphylococcus aureus and its Resistance to Methicillin among Specimens from Kirkuk Community. Annals of the Romanian Society for Cell Biology, 25(7), 461-473.

[19]. Al-Azawi, I. H. S. (2013). Antibiotic susceptibility pattern and mecA gene detection in methicillin resistance Staphylococcus aureus (MRSA) Isolated from burn and wound in Al-Diwaniya city. Journal of Babylon University/Pure and Applied Sciences, 3(21), 917-926.

[20]. Ibed, A. N., & Hamim, S. S. (2014). Molecular detection of methicillin resistant staphylococcus aureus isolated from burns infection in Al-nasiriyah city. World Journal of Pharmaceutical Sciences, 2(9),950-954

[21]. Tasnim, A., Shamsuzzaman, A. K. M., Alam, M. S., Ferdose, J., & Khan, F. A. (2020). Phenotypic and Genotypic Isolation of MRSA from Burn Wound Infected Patients at a Tertiary Care Hospital in Bangladesh. Bangladesh Journal of Infectious Diseases, 7(1), 12-17.

[22]. Kharre, A., Patel, V. K., Shukla, V., Gupta, R., Singh, A., & Bisen, J. (2020). Methicillin-resistant Staphylococcus aureus-prevalence and Antimicrobial Sensitivity Pattern among Burn Patients in a Tertiary Care Hospital in Vindhya Region. INTERNATIONAL JOURNAL OF SCIENTIFIC STUDY, 7(11), 102-107.

[23]. Clarridge III, J. E. (2004). Impact of 16S rRNA gene sequence analysis for identification of bacteria on clinical microbiology and infectious diseases. Clinical microbiology reviews, 17(4), 840-862.

[24]. Rasheed, N. A., Abdulrahman, R. F., & Hussein, N. R. (2022). Phylogenetic relatedness of methicillin-resistant Staphylococcus aureus isolates from the host community and Syrian refugees in Duhok Governorate based on 16S rRNA. IJID Regions, 4, 42-46.

[25]. Mehndiratta, P. L., & Bhalla, P. (2012). Typing of Methicillin resistant Staphylococcus aureus: a technical review. Indian journal of medical microbiology, 30(1), 16-23.

[26]. Trindade, P. A., Mcculloch, J. A., Oliveira, G. A. D., & Mamizuka, E. M. (2003). Molecular techniques for MRSA typing: current issues and perspectives. Brazilian Journal of Infectious Diseases, 7, 32-43.

[27]. Stranden, A., Frei, R., & Widmer, A. F. (2003). Molecular typing of methicillin-resistant Staphylococcus aureus: can PCR replace pulsed-field gel electrophoresis?. Journal of clinical microbiology, 41(7), 3181-3186.

[28]. Hussein, N. R., Alyas, A., Majeed, M., & Assafi, M. S. (2015). Prevalence rate and prevalent genotypes of ca-mrsa in kurdistan region: First report from iraq. International Journal of Pure and Applied Sciences and Technology, 27(1), 44.

[29]. Takeda, S., Yasunaka, K., Kono, K., & Arakawa, K. (2000). Methicillin-resistant Staphylococcus aureus (MRSA) isolated at Fukuoka University Hospital and hospitals and clinics in the Fukuoka city area. International journal of antimicrobial agents, 14(1), 39-43.

[30]. Alp, E., Klaassen, C. H., Doganay, M., Altoparlak, U., Aydin, K., Engin, A., ... & Voss, A. (2009). MRSA genotypes in Turkey: persistence over 10 years of a single clone of ST239. Journal of Infection, 58(6), 433-438.

[31]. Ho, W. Y., Choo, Q. C., & Chew, C. H. (2017). Predominance of three closely related methicillin-resistant Staphylococcus aureus clones carrying a unique ccrC-positive SCC mec type III and the emergence of spa t304 and t690 SCC mec type IV pvl+ MRSA Isolates in Kinta Valley, Malaysia. Microbial Drug Resistance, 23(2),215-223.

[32]. Aqel, A. A., Alzoubi, H. M., Vickers, A., Pichon, B., & Kearns, A. M. (2015). Molecular epidemiology of nasal isolates of methicillin-resistant Staphylococcus aureus from Jordan. Journal of Infection and Public Health, 8(1), 90-97.

[33]. Hussein, N. R., Assafi, M. S., & Ijaz, T. (2017). Methicillin-resistant Staphylococcus aureus nasal colonization amongst healthcare workers in Kurdistan Region, Iraq. Journal of global antimicrobial resistance, 9, 78-81.

[34]. Iyer, A., Kumosani, T., Azhar, E., Barbour, E., & Harakeh, S. (2014). High incidence rate of methicillin-resistant Staphylococcus aureus (MRSA) among healthcare workers in Saudi Arabia. The Journal of Infection in Developing Countries, 8(03), 372-378.