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Soltani Fard E, Roayaei Ardakani M, Motamedi H. Molecular Diversity of Methicillin-resistant Staphylococcus aureus Isolates Originated from Patients in Ahvaz Hospitals, Iran. JoMMID 2019; 7 (1 and 2) :19-28
URL: http://jommid.pasteur.ac.ir/article-1-181-en.html
Department of Biology, Faculty of Science, Shahid Chamran University of Ahvaz, Ahvaz, Iran
Abstract:   (4030 Views)
Introduction: Staphylococcus aureus is among the primary cause of hospitals and community-acquired infections. The emergence of methicillin-resistant S. aureus (MRSA) strains has resulted in the treatment failure of the infections caused by these bacteria. Hence, regional data on antibiotic resistance of S. aureus strains is necessary to adopt appropriate treatment regimens. This study aims to identify the diversities and their frequencies among MRSA isolates by molecular analysis of four genes. Methods: In a cross-sectional study, 100 S. aureus isolates from patients hospitalized in two hospitals of Ahvaz, Iran were collected and identified. The MRSA isolates were identified by phenotypic method and amplification of the mecA gene. The diversity of MRSA isolates was investigated by amplification of the coa, spa, aroA, and gap genes followed by RFLP analysis using the AluI, HindIII, TaqI and RsaI restriction enzymes. Results: In this study, we identified 50 MRSA isolates. Based on the analysis of coa gene, 8 types, spa gene 5 types and 17 subtypes, coa gene with AluI 13 types, and spa with HindIII 13 types were identified. Also, the RFLP analysis of gap gene with AluI revealed 3 types, and of aroA gene with TaqI and RsaI, 3 types and 2 subtypes, respectively.  Conclusion: Our PCR-RFLP analysis revealed that diversities are present among MRSA isolates originated from clinical samples and showed that this method is simple, reproducible, and cost-effective. 
Full-Text [PDF 647 kb]   (1787 Downloads)    
Type of Study: Original article | Subject: Epidemiologic studies including microbial genotyping, phenotyping and serotyping
Received: 2018/12/4 | Accepted: 2019/06/30 | Published: 2019/11/3

References
1. 1. Saei HD, Ahmadi M, Mardani K, Batavani R. Molecular typing of Staphylococcus aureus isolated from bovine mastitis based on polymorphism of the coagulase gene in the north west of Iran. Vet Microbiol. 2009; 137 (1): 202-6. [DOI:10.1016/j.vetmic.2009.01.001]
2. Gould IM, David MZ, Esposito S, Garau J, Lina G, Mazzei T, et al. New insights into meticillin-resistant Staphylococcus aureus (MRSA) pathogenesis, treatment and resistance. Int J Antimicrob Agents. 2012; 39 (2): 96-104. [DOI:10.1016/j.ijantimicag.2011.09.028]
3. Shakeri F, Ghaemi EA, Kochkaksaraei MB. Staphylococcus aureus typing by digestion of protein A coding gene using Bsp143I. Jundishapur J Microbiol. 2014; 7 (6). [DOI:10.5812/jjm.10320]
4. Hookey J, Edwards V, Cookson B, Richardson J. PCR-RFLP analysis of the coagulase gene of Staphylococcus aureus: application to the differentiation of epidemic and sporadic methicillin-resistant strains. J Hos Infect. 1999; 42 (3): 205-12. [DOI:10.1053/jhin.1999.0595]
5. Tenover FC, Arbeit R, Archer G, Biddle J, Byrne S, Goering R, et al. Comparison of traditional and molecular methods of typing isolates of Staphylococcus aureus. J Clin Microbiol. 1994; 32 (2): 407-15.
6. Mostafa SI. Molecular typing of methicilin resistant Staphylococcus aureus by spa gene polymorphism. Afr J Microbiol Res (AJMA). 2013; 7 (9): 755-9
7. Tambic A, Power E, Tambic T, Snur I, French G. Epidemiological analysis of methicillin-resistant Staphylococcus aureus in a Zagreb trauma hospital using a randomly amplified polymorphic DNA-typing method. Euro J Clin Microbiol Infect Dis. 1999; 18 (5): 335-40. [DOI:10.1007/PL00015015]
8. Talebi-Satlou R, Ahmadi M, Dastmalchi Saei H. Restriction fragment length polymorphism genotyping of human Staphylococcus aureus isolates from two hospitals in Urmia region of Iran using the coa gene. Jundishapur J Microbiol. 2012; 5 (2): 416-420. [DOI:10.5812/jjm.3522]
9. Zadoks R, Van Leeuwen W, Barkema H, Sampimon O, Verbrugh H, Schukken YH, et al. Application of Pulsed-Field Gel Electrophoresis and Binary Typing as Tools in Veterinary Clinical Microbiology and Molecular Epidemiologic Analysis of Bovine and Human Staphylococcus aureus Isolates. J Clin Microbiol. 2000; 38 (5): 1931-9.
10. Willems RJ, Hanage WP, Bessen DE, Feil EJ. Population biology of Gram-positive pathogens: high-risk clones for dissemination of antibiotic resistance. FEMS Microbiol Rev. 2011; 35 (5): 872-900. [DOI:10.1111/j.1574-6976.2011.00284.x]
11. Afrough P, Pourmand MR, Zeinalinia N, Yousefi M, Abdossamadi Z, Bagherzadeh et al. Molecular typing of clinical and nasal carriage isolates of staphylococcus aureus by spa gene patterns. J Mazandaran Univ Med Sci, 2012; 22 (94): 28-34.
12. Mehndiratta P, Bhalla P, Ahmed A, Sharma Y. Molecular typing of methicillin-resistant Staphylococcus aureus strains by PCR-RFLP of SPA gene: a reference laboratory perspective. Indian J Med Microbiol (IJMM). 2009; 27 (2): 116. [DOI:10.4103/0255-0857.45363]
13. Agius P, Kreiswirth B, Naidich S, Bennett KP. Typing Staphylococcus- aureus using the spa gene and novel distance measures. IEEE/ACM Trans Comput Biol Bioinform. 2007; 4 (4): 693-704. [DOI:10.1109/tcbb.2007.1053]
14. Baum C, Haslinger-Löffler B, Westh H, Boye K, Peters G, Neumann C, et al. Non-spa-typeable clinical Staphylococcus aureus strains are naturally occurring protein A mutants. J Clin Microbiol. 2009; 47 (11): 3624-9. [DOI:10.1128/JCM.00941-09]
15. Marcos JY, Soriano AC, Salazar MS, Moral CH, Ramos SS, Smeltzer MS, et al. Rapid Identification and Typing of Staphylococcus aureus by PCR-Restriction Fragment Length Polymorphism Analysis of the aroA Gene. J Clin Microbiol. 1999; 37 (3): 570-4.
16. Yugueros J, Temprano A, Berzal B, Sánchez Ma, Hernanz C, Luengo JMa, et al. Glyceraldehyde-3-Phosphate Dehydrogenase-Encoding Gene as a Useful Taxonomic Tool for Staphylococcusspp. J Clin Microbiol, 2000; 38 (12): 4351-5.
17. Brown DF, Edwards DI, Hawkey PM, Morrison D, Ridgway GL, Towner KJ, et al, Chemotherapy JWPotBSfA. Guidelines for the laboratory diagnosis and susceptibility testing of methicillin-resistant Staphylococcus aureus (MRSA). J Antimicrob Chemother. 2005; 56 (6): 1000-18. [DOI:10.1093/jac/dki372]
18. Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman J, Smith JA, et al. Short protocols in molecular biology. 1992.
19. McClure J, Conly JM, Lau G. Novel multiplex PCR assay for detection of the staphylococcal virulence marker Panton-Valentine leukocidin genes and simultaneous discrimination of methicillin-susceptible from resistant staphylococci. J Clin Microbiol. 2006; 44: 1141-4. [DOI:10.1128/JCM.44.3.1141-1144.2006]
20. Hookey JV, Richardson JF, Cookson BD. Molecular typing of Staphylococcus aureus based on PCR restriction fragment length polymorphism and DNA sequence analysis of the coagulase gene. J Clin Microbiol. 1998; 36 (4):1083-9.
21. Wichelhaus TA, Hunfeld K-P, Böddinghaus B, Kraiczy P, Schafer V, Brade V. Rapid molecular typing of methicillin-resistant Staphylococcus aureus by PCR-RFLP. Control Hosp Epidemiol. 2001; 22 (05): 294-8. [DOI:10.1086/501903]
22. Yugueros J, Temprano A, Sánchez Ma, Luengo JMa, Naharro G. Identification of Staphylococcus spp. by PCR-Restriction Fragment Length Polymorphism of gapGene. J Clin Microbiol. 2001; 39 (10): 3693-5. [DOI:10.1128/JCM.39.10.3693-3695.2001]
23. El-Huneidi W, Bdour S, Mahasneh A. Detection of enterotoxin genes seg, seh, sei, and sej and of a novel aroA genotype in Jordanian clinical isolates of Staphylococcus aureus. Diagn Microbiol Infect Dis. 2006; 56 (2): 127-32. [DOI:10.1016/j.diagmicrobio.2006.04.002]
24. Maleki SaA, Z. Comparison of the sensitivity of the results issued by two different anti-germ disc and E test for oxa‌cillin and vancomycin antibiotics. Islamic Azad Univ J Med Sci. 2007; 16 (4): 211-15.
25. Afrough P, Pourmand MR, Sarajian AA, Saki M, Saremy S. Molecular investigation of Staphylococcus aureus, coa and spa genes in Ahvaz hospitals, staff nose compared with patients clinical samples. JundiShapur J Microbiol. 2013; 6 (4). [DOI:10.5812/jjm.5377]
26. Zeinali E, Moniri R, Safari M, Mousavi SGA. Molecular characterization and SCCmec typing in meticillin-resistant Staphylococcus aureus isolated from clinical samples. J Kaums (FEYZ). 2011; 14 (4): 439-46.
27. Khosravi AD, Hoveizavi H, Farshadzadeh Z. The prevalence of genes encoding leukocidins in Staphylococcus aureus strains resistant and sensitive to methicillin isolated from burn patients in Taleghani Hospital, Ahvaz, Iran. Burns. 2012; 38 (2): 247-51. [DOI:10.1016/j.burns.2011.08.002]
28. Japoni A, Jamalidoust M, Farshad S, Ziyaeyan M, Alborzi A, Japoni S, et al. Characterization of SCCmec types and antibacterial susceptibility patterns of methicillin-resistant Staphylococcus aureus in Southern Iran. Japanese J Infect Dis (JJID). 2011; 64 (1): 28-33.
29. Lee BY, Singh A, David MZ, Bartsch SM, Slayton RB, Huang SS, et al. The economic burden of community‐associated methicillin‐resistant Staphylococcus aureus (CA‐MRSA). Clin Microbial Infect. 2013; 19 (6): 528-36. [DOI:10.1111/j.1469-0691.2012.03914.x]
30. Ishino K, Tsuchizaki N, Ishikawa J, Hotta K. Usefulness of PCR-restriction fragment length polymorphism typing of the coagulase gene to discriminate arbekacin-resistant methicillin-resistant Staphylococcus aureus strains. J Clin Microbiol. 2007; 45 (2): 607-9. [DOI:10.1128/JCM.02099-06]
31. Janwithayanuchit I, Ngam-Ululert S, Paungmoung P, Rangsipanuratn W. Epidemiologic study of methicillin-resistant Staphylococcus aureus by coagulase gene polymorphism. Science Asia. 2006; 32: 127-32. [DOI:10.2306/scienceasia1513-1874.2006.32.127]
32. Omar NY, Ali HAS, Harfoush RAH, El Khayat EH. Molecular typing of methicillin resistant staphylococcus aureus clinical isolates on the basis of protein A and coagulase gene polymorphisms. Int J Microbiol. 2014; 2014. [DOI:10.1155/2014/650328]
33. da Silva ER, da Silva N. Coagulase gene typing of Staphylococcus aureus isolated from cows with mastitis in southeastern Brazil. Can J Vet Res. 2005; 69 (4): 260.
34. Himabindu M, Muthamilselvan DS, Bishi DK, Verma RS. Molecular analysis of coagulase gene polymorphism in clinical isolates of methicillin resistant Staphylococcus aureus by restriction fragment length polymorphism based genotyping. Am J Infect Dis. 2009; 5: 170-6. [DOI:10.3844/ajidsp.2009.163.169]
35. Schmitz FJ MS, HV. Tichy, et al. Typing of methicillin-resistant Staphylococcus aureus isolates 1from Dusseldorf by six genotypic methods. J Med Microbiol. 1998; 47 (4): 341-51. [DOI:10.1099/00222615-47-4-341]
36. Montesinos I, Salido E, Delgado T, Cuervo M, Sierra A. Epidemiologic genotyping of methicillin-resistant Staphylococcus aureus by pulsed-field gel electrophoresis at a university hospital and comparison with antibiotyping and protein A and coagulase gene polymorphisms. J Clin Microbiol. 2002; 40 (6): 2119-25. [DOI:10.1128/JCM.40.6.2119-2125.2002]
37. Adesida SA, Likhoshvay Y, Eisner W, Coker AO, Abioye OA, Ogunsola FT. Repeats in the 3'region of the protein A gene is unique in a strain of Staphylococcus aureus recovered from wound infections in Lagos, Nigeria. Afr J Biotechnol. 2006; 5 (20): 1858.
38. Soltan-Dallal M, Salehipour Z, Mehrabadi JF. Molecular epidemiology of Staphylococcus aureus in food samples based on the protein A gene polymorphic region DNA sequence. Can J Microbiol. 2010; 56 (1): 18-21. [DOI:10.1139/W09-111]
39. Mahmoudi H, Arabestani MR, Mousavi SF, Ghafel S, Alikhani MY. Study of polymorphism spa gene (encoding protein A) of Staphylococcus aureus in clinical isolates and nasal carriers. Tehran Univ Med J. 2015; 73 (1): 24-30.
40. Talebi-Satlou R, Ahmadi M, Ghavam F, Saei HD. Use of 5-Enolpyruvylshikmate-3-Phosphate synthase encoding gene for typing of Staphylococcus aureus isolated from skin and urinary tract infections of human. Iran J Basic Med Sci (ijbms). 2011; 15(4): 975-982.

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Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.