Volume 11, Issue 6 (Nov - Dec 2017)                   mljgoums 2017, 11(6): 12-17 | Back to browse issues page


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Hakimi Alni R, Mohammadzadeh A, Mahmoodi P, Alikhani M Y. Genotypic Analysis of Staphylococcus aureus Coagulase Gene using PCR-RFLP analysis . mljgoums 2017; 11 (6) :12-17
URL: http://mlj.goums.ac.ir/article-1-1039-en.html
1- Department of Pathobiology, Faculty of Veterinary Sciences, Bu-Ali Sina University, Hamedan, Ira
2- Department of Pathobiology, Faculty of Veterinary Sciences, Bu-Ali Sina University, Hamedan, Ira , mohammadzadeh@basu.ac.ir
3- Department of Pathobiology, Faculty of Veterinary Science, Bu-Ali Sina University, Hamedan, Iran
4- Department of Microbiology, Faculty of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran
Abstract:   (11286 Views)
ABSTRACT
          Background and Objectives: Determining the genetic relationship between S. aureus isolates is important for epidemiological surveillance and control of infections caused by this bacterium. The present study was conducted to determine polymorphisms of coagulase gene (coa) among S. aureus isolates from pastry and cheese samples using restriction fragment length polymorphism (RFLP) analysis.
         Methods: Overall, 65 S. aureus isolated from pastry (n=45) and cheese (n=20) samples were examined for the coa gene by polymerase chain reaction (PCR). PCR products were digested with AluI enzyme and the products were assessed using gel electrophoresis.
          Results: Except for two isolates, all isolates were positive in coa-PCR and produced four different PCR products, with molecular sizes ranging from 570 to 970 bp. Overall; five distinct RFLP patterns were detected (I-V). Although pattern types I and III were present in isolates from both samples, types I and IV were mainly present in isolates from cheese and pastry samples, respectively.
        Conclusion: PCR-RFLP analysis of the coa gene indicates that S. aureus isolates from pastry and cheese samples may be originated from different sources. However, as one pattern type was predominant in each group, it can be concluded that majority of the isolates may have the same origin.
          Keywords: Staphylococcus aureus, PCR-RFLP, Coagulase, Pastry, Cheese.
Full-Text [PDF 546 kb]   (1588 Downloads)    
Research Article: Original Paper |
Received: 2018/01/22 | Accepted: 2018/01/22 | Published: 2018/01/22 | ePublished: 2018/01/22

References
1. Normanno G, Firinu A, Virgilio S, Mula G, Dambrosio A, Poggiu A, et al. Coagulase-positive Staphylococci and Staphylococcus aureus in food products marketed in Italy. Int J Food Microbiol. 2005; 98(1): 73-9. [DOI:10.1016/j.ijfoodmicro.2004.05.008]
2. Bendahou A, Lebbadi M, Ennanei L, Essadqui FZ, Abid M. Characterization of Staphylococcus species isolated from raw milk and milk products (lben and jben) in North Morocco. J Infect Dev Ctries. 2008; 2(03): 218-25.
3. Normanno G, La Salandra G, Dambrosio A, Quaglia N, Corrente M, Parisi A, et al. Occurrence, characterization and antimicrobial resistance of enterotoxigenic Staphylococcus aureus isolated from meat and dairy products. Int J Food Microbiol. 2007; 115(3): 290-6. DOI:10.1016/j.ijfoodmicro.2006.10.049 [DOI:10.1016/j.ijfoodmicro.2006.10.049]
4. Mehndiratta P, Bhalla P. Typing of Methicillin resistant Staphylococcus aureus: a technical review. Indian J Med Microbiol. 2012; 30(1):16-23. doi: 10.4103/0255-0857.93015. [DOI:10.4103/0255-0857.93015]
5. Laplana LM, Cepero MPG, Ruiz J, Zolezzi PC, Calvo MCR, Erazo MC, et al. Molecular typing of Staphylococcus aureus clinical isolates by pulsed-field gel electrophoresis, staphylococcal cassette chromosome mec type determination and dissemination of antibiotic resistance genes. Int J Antimicrob Agents. 2007; 30(6): 505-13. DOI:10.1016/j.ijantimicag.2007.06.020. [DOI:10.1016/j.ijantimicag.2007.06.020]
6. Larsen J, Enright MC, Godoy D, Spratt BG, Larsen AR, Skov RL. Multilocus sequence typing scheme for Staphylococcus aureus: revision of the gmk locus. J Clin Microbiol. 2012; 50(7): 2538-9. doi: 10.1128/JCM.00290-12. [DOI:10.1128/JCM.00290-12]
7. 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.
8. 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.
9. Alni RH, Mohammadzadeh A, Mahmoodi P, Alikhani MY. RAPD-PCR analysis of Staphylococcus aureus strains isolated from different sources. Comp Clin Path. 2017; 26(4): 823-830. [DOI:10.1007/s00580-017-2453-z]
10. Friedrich R, Panizzi P, Fuentes-Prior P, Richter K, Verhamme I, Anderson PJ, et al. Staphylocoagulase is a prototype for the mechanism of cofactor-induced zymogen activation. Nature. 2003; 425(6957): 535-9. DOI:10.1038/nature01962. [DOI:10.1038/nature01962]
11. Wichelhaus TA, Hunfeld K-P, Böddinghaus B, Kraiczy P, Schafer V, Brade V. Rapid molecular typing of methicillin-resistant Staphylococcus aureus by PCR-RFLP. Infect Control Hosp Epidemiol. 2001; 22(05): 294-8. [DOI:10.1086/501903]
12. Van Belkum A, Scherer S, Van Alphen L, Verbrugh H. Short-sequence DNA repeats in prokaryotic genomes. Microbiology and Molecular Biology Reviews. 19981; 62(2): 275-93.
13. Wilson K. Preparation of genomic DNA from bacteria. Curr Protoc Mol Biol. 2001; 2(4): 1-2. [DOI:10.1002/0471142727.mb0204s56]
14. Goh SH, Byrne S, Zhang J, Chow A. Molecular typing of Staphylococcus aureus on the basis of coagulase gene polymorphisms. J Clin Microbiol. 1992; 30(7): 1642-5.
15. Karahan M, Cetinkaya B. Coagulase gene polymorphisms detected by PCR in Staphylococcus aureus isolated from subclinical bovine mastitis in Turkey. Vet J. 2007; 174(2): 428-31. DOI:10.1016/j.tvjl.2006.05.016. [DOI:10.1016/j.tvjl.2006.05.016]
16. Aarestrup FM, Dangler C, Sordillo L. Prevalence of coagulase gene polymorphism in Staphylococcus aureus isolates causing bovine mastitis. Can J Vet Res. 1995; 59(2): 124-128.
17. Momtaz H, Tajbakhsh E, Rahimi E, Momeni M. Coagulase gene polymorphism of Staphylococcus aureus isolated from clinical and sub-clinical bovine mastitis in Isfahan and Chaharmahal va Bakhtiari provinces of Iran. Comp Clin Path. 2011; 20(5): 519-22. [DOI:10.1007/s00580-010-1029-y]
18. Khan A, Hussain R, Javed MT, Mahmood F. Molecular analysis of virulent genes (coa and spa) of Staphylococcus aureus involved in natural cases of bovine mastitis. Pak J Agric Sci. 2013; 50: 739-43.
19. Tiwari HK, Sapkota D, Gaur A, Mathuria JP, Singh A, Sen MR. Molecular typing of clinical Staphylococcus aureus isolates from northern India using coagulase gene PCR-RFLP. Southeast Asian J Trop Med Public Health. 2008; 39(3): 467-73.
20. Mahmoudi H, Arabestani MR, Mousavi SF, Alikhani MY. Molecular analysis of the coagulase gene in clinical and nasal carrier isolates of methicillin-resistant Staphylococcus aureus by restriction fragment length polymorphism. J Glob Antimicrob Resist. 2017; 8: 41-45. doi: 10.1016/j.jgar.2016.10.007. [DOI:10.1016/j.jgar.2016.10.007]
21. Reinoso E, El-Sayed A, Lämmler C, Bogni C, Zschöck M. Genotyping of Staphylococcus aureus isolated from humans, bovine subclinical mastitis and food samples in Argentina. Microbiol Res. 2008; 163(3): 314-22. [DOI:10.1016/j.micres.2006.05.013]
22. Scherrer D, Corti S, Muehlherr J, Zweifel C, Stephan R. Phenotypic and genotypic characteristics of Staphylococcus aureus isolates from raw bulk-tank milk samples of goats and sheep. Vet Microbiol. 2004; 101(2): 101-7. [DOI:10.1016/j.vetmic.2004.03.016]
23. 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. J J Microbiol. 2012; 2(3): 416-20.[Persian]. [DOI:10.5812/jjm.3522]
24. aureus clinical isolates on the basis of protein A and coagulase gene polymorphisms. Int J Microbiol. 2014; 15: 1-11. doi: 10.1155/2014/650328. [DOI:10.1155/2014/650328]
25. Lee JH. Methicillin (oxacillin)-resistant Staphylococcus aureus strains isolated from major food animals and their potential transmission to humans. Appl Environ Microbiol. 2003; 69(11): 6489-94. [DOI:10.1128/AEM.69.11.6489-6494.2003]
26. Nnachi AU, Emele FE, Ukaegbu CO, Agah MV, Udu-Ibiam OE, Chukwu OS, et al. Prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in raw meat and meat handlers in Onitsha, Nigeria. Europ J Prev Med. 2014; 2: 9-15. doi: 10.3390/ijerph13100949. [DOI:10.3390/ijerph13100949]

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