Abstract
A total of 100 Staphylococcus aureus strains were isolated from 1,047 food samples. In addition to biotyping all isolates, the occurrence of the tst gene and protein were evaluated by PCR and the RPLA test, respectively. Moreover, polymorphism of the X-region of the protein A gene was analyzed by PCR-RFLP. TSST1 was detected in 12 strains and production of TSST1 in all strains was confirmed by RPLA assay. It was noteworthy that 66.7% of TSST1-producing S. aureus strains belonged to the human biotype. The result of genotyping amplification of the spa gene displayed size polymorphisms and revealed seven different clusters ranging from 1,200 bp to 1,600 bp. Digestion of amplicons with HindIII and HaeII resulted in six distinct patterns for each of them. An amplicon of 1,450 bp in size was the predominant type of Spa-PCR product. The accuracy of the Spa typing system by PCR-RFLP was 100%, thus confirming this method as a reliable tool in routine epidemiological investigation for S. aureus.
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Adesiyun AA, Lenz W, Schaal KP (1992) Production of Toxic Shock Syndrome Toxin-1 (TSST-1) by Staphylococcus aureus strains isolated from humans, animals and foods in Nigeria. Microbiologica 15:125–133
Akineden O, Annemuller C, Hassan AA, Lammler C, Wolter W, Zschock M (2001) Toxin genes and other characteristics of Staphylococcus aureus isolates from milk of cows with mastitis. Clin Vac Immunol 8:959–964
Annemüller C, Lammler C, Zschock M (1999) Genotyping of Staphylococcus aureus isolated from bovine mastitis. Vet Microbiol 69:217–224
Atanassova V, Meindl A, Ring C (2001) Prevalence of Staphylococcus aureus and Staphylococcal enterotoxins in raw pork and uncooked smoked ham—a comparison of classical culturing detection and RFLP-PCR. Int J Food Microbiol 68:105–113
Capita R, Alonso-Calleja C, Garcia-Fernandez MC, Moreno B (2002) Characterization of Staphylococcus aureus isolated from poultry meat in Spain. Poultry Sci 81:414–421
Casciano R, Alberghini L, Peccio A, Serraino A, Rosmini R (2003) Typing of Staphylococcus aureus isolates from raw milk. Vet Res Commun 27:289–291
Chapaval L, Moon DH, Gomes JE, Duarte FR, Tsai SM (2006) Use of PCR to detect classical enterotoxins genes (ent) and toxic shock syndrome toxin-1 gene (tst) in Staphylococcus aureus isolated from crude milk and determination of toxin productivities of S. aureus isolates harboring these genes. Arq Inst Biol Sao Paulo 73:165–169
Devriese LA (1984) A simplified system for biotyping Staphylococcus aureus strains isolated from different animal species. J Appl Microbiol 56:215–220
El-Ghodban A, Ghenghesh KS, Marialigeti K, Esahli H, Tawil A (2006) PCR detection of toxic shock syndrome toxin of Staphylococcus aureus from Tripoli. Libya. J Med Microbiol 55:179–182
Fueyo JM, Mendoza MC, Martin MC (2005a) Enterotoxins and toxic shock syndrome toxin in Staphylococcus aureus recovered from human nasal carriers and manually handled foods: epidemiological and genetic findings. Microbes Infect 7:187–194
Fueyo JM, Mendoza MC, Rodicio MR, Muniz J, Alvarez MA, Martin MC (2005b) Cytotoxin and pyrogenic toxin superantigen gene profiles of Staphylococcus aureus associated with subclinical mastitis in dairy cows and relationships with macrorestriction genomic profiles. J Clin Microbiol 43:1278–1284
Hallin M, Deplano A, Denis O, De Mendonça R, De Ryck R, Struelens MJ (2007) Validation of pulsed-field gel electrophoresis and spa typing for long-term, nationwide epidemiological surveillance studies of Staphylococcus aureus infections. J Clin Microbiol 45:127–133
Hata E, Katsuda K, Kobayashi H, Ogawa T, Endo T, Eguchi M (2005) Characteristics and epidemiological genotyping of Staphylococcus aureus isolates from bovine mastitic milk in Hokkaido, Japan. J Vet Med Sci 68:165–170
Hwang SY, Kim SH, Jang EJ, Kwon NH, Park YK, Koo HC, Jung WK, Kim JM, Park YH (2007) Novel Multiplex PCR for the detection of the Staphylococcus aureus superantigen and its application to raw meat isolates in Korea. Int J Food Microbiol 117:99–105
Jakubczak A, Szweda P, Lukaszewska K, Kur J (2007) Molecular typing of Staphylococcus aureus isolated from cows with mastitis in the east of poland on the basis of polymorphism of genes coding protein A and coagulase. Pol J Vet Sci 10:199–205
Jorgensen HJ, Mork T, Hogasen HR, Rorvik LM (2005) Enterotoxigenic Staphylococcus aureus in bulk milk in Norway. J Appl Microbiol 99:158–166
Katsuda K, Hata E, Kobayashi H, Kohmoto M, Kawashima K, Tsunemitsu H, Eguchi M (2005) Molecular typing of Staphylococcus aureus isolated from bovine mastitic milk on the basis of toxin genes and coagulase gene polymorphisms. Vet Microbiol 105:301–305
Kérouanton A, Hennekinne JA, Letertre C, Petit L, Chesneau O, Brisabois A, De Buyser ML (2007) Characterization of Staphylococcus aureus strains associated with food poisoning outbreaks in France. Int J Food Microbiol 115:369–375
Kitai S, Shimizu A, Kawano J, Sato E, Nakano C, Kitagawa H, Fujio K, Matsumura K, Yasuda R, Inamoto T (2005) Prevalence and characterization of Staphylococcus aureus and enterotoxigenic Staphylococcus aureus in retail raw chicken meat throughout Japan. J Vet Med Sci 67:269–274
Jung HJ, Cho JI, Park SH, Ha SD, Lee KH, Kim CH, Song ES, Chung DH, Kim MG, Kim KY, Kim KS (2005) Genotypic and phenotypic characteristics of Staphylococcus aureus Isolates from lettuces and raw milk. Korean J Food Sci Technol 37:134–141
Manfreda G, Mioni R, Cesare A (2005) Surveillance and Characterization of enterotoxigenic staphylococci in foods of animal origin collected in the Veneto Region. Vet Res Comm 29:331–333
Nagase N, Shimizu A, Kawano J, Yamashita K, Yoshimura H, Ishimaru M, Kojima A (2002) Characterization of Staphylococcus aureus strains isolated from bovine mastitis in Japan. J Vet Med Sci 64:1169–1172
Normanno G, Firinu A, Virgilio S, Mula G, Dambrosio A, Poggiu A, Decastelli L, Mioni R, Scuota S, Bolzoni G (2005) Coagulase-positive staphylococci and Staphylococcus aureus in food products marketed in Italy. Int J Food Microbiol 98:73–79
Normanno G, La Salandra G, Dambrosio A, Quaglia NC, Corrente M, Parisi A, Santagada G, Firinu A, Crisetti E, Celano GV (2007) Occurrence, Characterization and antimicrobial resistance of enterotoxigenic Staphylococcus aureus isolated from meat and dairy products. Int J Food Microbiol 115:290–296
Oh SUK, Lee N, Cho Y, Shin D, Choi SY, Koo M (2007) Occurrence of toxigenic Staphylococcus aureus in ready-to-eat food in Korea. J Food Protect 70:1153–1158
Rodríguez-Calleja JM, García-López I, Santos JA, Otero A, García-Lopez M-L (2006) Molecular and phenotypic typing of Staphylococcus aureus isolates from rabbit meat. Res Microbiol 157:496–502
Strommenger B, Braulke C, Heuck D, Schmidt C, Pasemann B, Nübel U, Witte W (2008) spa typing of Staphylococcus aureus as a frontline tool in epidemiological typing. J Clin Microbiol 46:574–581
Wichelhaus TA, Hunfeld KP, Boddinghaus B, Kraiczy P, Schafer V, Brade V (2001) Rapid molecular typing of methicillin resistant Staphylococcus aureus by PCR-RFLP. Infect Cont Hospital Epidemiol 22:294–298
Wu F, Della-Latta P (2002) Molecular typing strategies. Semin Perinatol 26:357–366
Zschock M, Botzler D, Blocher S, Sommerhouser J, Hamann HP (2000) Detection of genes for enterotoxins (ent) and toxic shock syndrome toxin-1 (tst) in mammary isolates of Staphylococcus aureus by polymerase-chain-reaction. Int Dairy J 10:569–574
Acknowledgments
We would like to thank S. Agha Amin for kindly helping in samples collection; A. Azimian (PhD student of microbiology, Isfahan University of Medical Science) for preparing reference strains; I. Ashrafi (Bacteriology Laboratory of the Veterinary Medicine Faculty, Tehran University of Medical Science) for kindly helping with biotyping, and M. Shokuhi (Physiology Research Centre, Kerman University of Medical Science) for helping us with data analysis.
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Soltan Dallal, M.M., Salehipour, Z., Eshraghi, S. et al. Occurrence and molecular characterization of Staphylococcus aureus strains isolated from meat and dairy products by PCR-RFLP. Ann Microbiol 60, 189–196 (2010). https://doi.org/10.1007/s13213-010-0025-4
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DOI: https://doi.org/10.1007/s13213-010-0025-4