Cemil DEMİR, Nizami DURAN, Burçin ÖZER, Özkan ASLANTAŞ, Sabahattin OCAK

Investigation of toxin genes in Staphylococcus aureus strains isolated in Mustafa Kemal University Hospital

The aim of this study was to investigate the presence of genes encoding staphylococcal enterotoxins (SEs), exfoliative toxins (ETAs, ETBs), and toxic shock syndrome toxin-1 (TSST-1) by polymerase chain reaction (PCR) in Staphylococcus aureus strains isolated from various clinical samples from the Mustafa Kemal University Hospital. In addition, PCR-based restriction fragment length polymorphism (RFLP) analysis of the coa gene was employed to genotype the isolates. Materials and methods: A total of 120 S. aureus strains isolated from various clinical samples (blood, wounds, urine, conjuctival swabs, and tracheal aspirate) over a 1 year period, 2007-2008, were used in this study. Results: Almost 65.8% of the isolates possessed at least one toxin gene. The genes most frequently found were seg-sei (40.8%), followed by sea (30%) and eta (19.2%). Overall, 35 toxin genotypes were observed, among which the genotypes seg-sei, sea-seg-sei, and sea-see predominated at the rate of 8.3%, 5.8%, and 5%, respectively. Four coagulase genotype patterns were observed, with molecular sizes ranging from 570 to 970 bp. Coa-based RFLP analysis revealed 7 different patterns using AluI. Conclusion: Our results have revealed that toxin genes were very prevalent among S. aureus isolates, and the toxigenic isolates were independent of the genotypes obtained by PCR-RFLP of the coa gene (P > 0.05).

Anahtar Kelimeler:

Key words: Staphylococcus aureus, toxin genes, PCR-RFLP, coagulase

Investigation of toxin genes in Staphylococcus aureus strains isolated in Mustafa Kemal University Hospital

Keywords:

Key words: Staphylococcus aureus, toxin genes, PCR-RFLP, coagulase,

PDF

___

Balaban N, Rasooly A. Staphylococcal enterotoxins. Int J Food Microbiol 2000; 61: 1-10.
Kotb M. Bacterial pyrogenic exotoxins as superantigens. Clin Microbiol 1995; 8: 411-426.
Lina G, Bohach GA, Nair SP, Hiramatsu K, Jouvin-Marche E, Mariuzza R. Standard nomenclature for the superantigens expressed by Staphylococcus. J Infect Dis 2004; 189: 2334- 2336.
Omoe K, Imanishi K, Hu DL, Kato H, Fugane Y, Abe Y et al. Characterization of novel staphylococcal enterotoxin-like toxin type P. Infect Immun 2005; 73: 5540–5546.
Omoe K, Hu DL, Takahashi-Omoe H, Nakane A, Shinagawa K. Identifi cation and characterization of a new staphylococcal enterotoxin-related putative toxin encoded by two kinds of plasmids. Infect Immun 2003; 71: 6088- 6094.
Leterte C, Perelle S, Dilaser F, Fach P. Identifi cation of a new putative enterotoxin SEU encoded by the egc cluster of Staphylococcus aureus. J Appl Microbiol 2003; 95: 38-43.
Orwin PM, Fitzgerald JR, Leung DY, Gutierrez JA. Bohach GA, Schlievert PM. Characterization of Staphylococcus aureus enterotoxin L. J Clin Microbiol 2003; 71: 2916-2919.
Omoe K, Ishikawa M, Shimoda Y, Hu DL, Ueda S, Shinagawa K. Detection of seg, seh, and sei genes in Staphylococcus aureus isolates and determination of the enterotoxin productivities of S. aureus isolates harboring seg, seh, or sei genes. J Clin Microbiol 2002; 40: 857-862.
Yarwood JM, McCormick JK, Paustian ML, Orwin PM, Kapur V, Schlievert PM. Characterization and expression analysis of Staphylococcus aureus pathogenicity island 3: implications for the evolution of staphylococcal pathogenicity island. J Biol Chem 2000; 277: 13138-13147.
Orwin PM, Leung DY, Donahue HL, Novick RP, Schlievert PM. Biochemical and biological properties of staphylococcal enterotoxin K. Infect Immun 2001; 69: 360- 366.
Jarraud S, Peyrat MA, Lim A, Tristan A, Bes M, Mougel C et al. egc, a highly prevalent operon of enterotoxin gene, forms a putative nursery of superantigens in Staphylococcus aureus. J Immunol 2001; 166: 669-677.
Su YC, Wong AC. Identifi cation and purifi cation of a new staphylococcal enterotoxin, H. Appl Environ Microbiol 1995; 61: 1438-1443.
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: 407-415.
Goh SH, Bryne SK, Zhang JL, Chow AW. Molecular typing of Staphylococcus aureus on the basis of coagulase gene polymorphisms. J Clin Microbiol 1992; 30: 1642-1645.
Hesselbarth J, Schwarz S. Comparative ribotyping of Staphylococcus intermedius from dogs, pigeons, horses and mink. Vet Microbiol 1995; 45: 11-17.
Mehrotra M, Wang G, Johnson WM. Multiplex PCR for detection of genes for Staphylococcus aureus enterotoxins, exfoliative toxins, toxic shock syndrome toxin-1, and methicillin resistance. J Clin Microbiol 2000; 38: 1032- 1035.
Monday SR, Bohach GA. Use of multiplex PCR to detect classical and newly described pyrogenic toxin genes in staphylococcal isolates. J Clin Microbiol 1999; 37: 3411- 3414.
Becker K, Friedrich AW, Lubritz G, Weilert M, Peters G, Von Eiff C. Prevalence of genes encoding pyrogenic toxin superantigens and exfoliative toxins among strains of Staphylococcus aureus isolated from blood and nasal specimens. J Clin Microbiol 2003; 41:1434-1439.
Naff a RG, Bdour SM, Migdadi HM, Shehabi AA. Enterotoxicity and genetic variation among clinical Staphylococcus aureus isolates in Jordan. J Med Microbiol 2006; 55: 183-187.
Chiang YC, Liao WW, Fan CM, Pai WY, Chiou CS, Tsen HY. PCR detection of Staphylococcal enterotoxins (SEs) N, O, P, Q, R, U, and survey of SE types in Staphylococcus aureus isolates from food-poisoning cases in Taiwan. Int J Food Microbiol 2008; 121: 66-73.
Tekeli A., Koyuncu E, Dolapçı İ, Akan ÖA. Karahan, ZC. Molecular characteristics of methicillin-resistant Staphylococcus aureus strains isolated from blood cultures between 2002-2005 in Ankara University Hospital. Mikrobiyol Bul 2009; 43: 1-10.
Fueyo JM, Martín MC, González-Hevia MA, Mendoza MC. Enterotoxin production and DNA fi ngerprinting in Staphylococcus aureus isolated from human and food samples. Relations between genetic types and enterotoxins. Int J Food Microbiol 2001; 20: 139-145.
Nashev D, Toshkova K, Salasia SI, Hassan AA, Lämmler C, Zschöck M. Distribution of virulence genes of Staphylococcus aureus isolated from stable nasal carriers. FEMS Microbiol Lett 2004; 233: 45-52.
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; 56:127-132.
Zhang S, Iandolo JJ, Stewart GC. Th e enterotoxin D plasmid of Staphylococcus aureus encodes a second enterotoxin determinant (sej). FEMS Microbiol Lett 1998; 168: 227-233.
Holtfreter S, Grumann D, Schmudde M, Nguyen HTT, Eichler P, Strommenger B, et al. Clonal distribution of superantigen genes in clinical Staphylococcus aureus isolates. J Clin Microbiol 2007; 45: 2669-2680.
Dancer SJ, Noble WC. Nasal, axillary, and perineal carriage of Staphylococcus aureus among women: Identifi cation of strains producing epidermolytic toxin. J Clin Pathol 1991; 44: 681-684.
Baba T, Takeuchi F, Kuroda M, Yuzawa H, Aoki K, Oguchi A et al. Genome and virulence determinants of high virulence community-acquired MRSA. Lancet 2002; 359: 1819-1827.