In the European Union, Staphylococcus aureus (S. aureus) is an important pathogenic agent of food intoxications (Denayer et al., 2017). S. aureus strains have several virulence factors such as lipases, thermonuclease, hyaluronidase, and haemolysins; however, the major virulence factors are the heat-stable enterotoxins that cause staphylococcal food poisoning (SFP) (Ortega et al., 2010). The onset of SFP symptoms is very rapid in contrast to other gastrointestinal infections, generally a short time after ingestion of the contaminated food (Mossong et al., 2015). In outbreaks, the symptoms typically include diarrhoea (89%), vomiting (87%), and abdominal cramps (72%); however, fever (9%) is rarely reported. Usually, pork (41% of pork dishes were ham) or poultry dishes (41% freshly prepared poultry dishes) are implicated (55%) in S. aureus outbreaks (Bennett et al., 2013).
Coagulase-positive Staphylococcus species have several defence mechanisms like immunoglobulin binding proteins - including staphylococcal protein A (SpA) - that assist the evasion of the host immune system (Foster, 2005). Protein A is produced by numerous species of Staphylococcus, and is a highly potent virulence as well as immune evasion factor (Balachandran et al., 2018). The 40-60 kDa cell-wall protein is secreted during the exponential growth phase and is encoded by the spa gene (Björk & Sjöquist, 1972). The synthesised protein is able to bind to the Fc and F (ab’) 2 regions of immunoglobulins (Forsgren, 1970) and thus may interfere with the immunising ability of vaccines that depend on the production of opsonophagocytic antibodies, because these inhibit the functions of B-cells (Daum & Spellberg, 2012).
The spa-typing of S. aureus isolates has two major aims: to analyse genetic microvariation in outbreak investigations and to analyse genetic macrovariation in population- and phylogenetic-based assaying (Hallin et al., 2009). A variable repeat region (spa) in the gene allows a reliable and rapid method to discriminate S. aureus isolates in outbreaks from those presumed to be epidemiologically unrelated (Koreen et al., 2004). Furthermore, the epidemiological definition makes it possible to compare the outbreaks occurring in clinical cases and animal husbandry. Given that spa-typing involves the sequencing of only one gene, it provides significant advantages in terms of time-to-result, standardisation, ease of use, and reproducibility as compared to other molecular biology techniques (Agius et al., 2007).
Matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI- TOF MS) allows the identification of microorganisms isolated from food or clinical cases by a low-cost, rapid, easy, high-throughput, and efficient identification technique (Croxatto et al., 2012). Using standardised and developed procedures, MALDI-TOF MS devices have revolutionised the identification at the species level of most Gram-positive and Gramnegative bacteria (Barnini et al., 2015). The present study aims to discriminate and perform molecular characterisation of foodborne coagulase-positive staphylococci typed molecular techniques after identification by MALDI-TOF MS.
Agius, P., Kreiswirth, B., Naidich, S. & Bennett, K. (2007): Typing Staphylococcus aureus using the spa gene and novel distance measures. IEEE/ACM T. Comput.Bi., 4, 693-704.
Argudin, M.A., Mendoza, M.C. & Rodicio, M.R. (2010): Food poisoning and Staphylococcus aureus enterotoxins. Toxins, 2(7), 1751-1773.
Balachandran, M., Bemis, D.A. & Kania, S.A. (2018): Expression and function of protein A in Staphylococcus pseudintermedius. Virulence, 9, 390-401.
Barnini, S., Ghelardi, E., Brucculeri, V., Morici, P. & Lupetti, A. (2015): Rapid and reliable identification of Gram-negative bacteria and Gram-positive cocci by deposition of bacteria harvested from blood cultures onto the MALDI-TOF plate. BMC Microbiol., 124, 1-8.
Bennett, S.D., Walsh, K.A. & Gould, L.H. (2013): Foodborne disease outbreaks caused by Bacillus cereus, Clostridium perfringens, and Staphylococcus aureus - United States, 1998-2008. Clin. Infect. Dis., 57, 425-433.
Bianchi, D.M., Gallina, S., Bellio, A., Chiesa, F., Civera, T. & Decastelli, L. (2014): Enterotoxin gene profiles of Staphylococcus aureus isolated from milk and dairy products in Italy. Lett. Appl. Microbiol., 58(2), 190-196.
Björk, I.P. & Sjöquist, J. (1972): Some physicochemical properties of protein A from Staphylococcus aureus. Eur. J. Biochem., 29, 579-584.
Croxatto, A., Prod’hom, G. & Greub, G. (2012): Applications of MALDI-TOF mass spectrometry in clinical diagnostic microbiology. FEMS Microbiol. Rev., 36, 380-407.
Denayer, S., Delbrassinne,L., Nia, Y. & Botteldoorn, N. (2017): Food-borne outbreak investigation and molecular typing: high diversity of Staphylococcus aureus strains and importance of toxin detection. Toxins, 9, 1-13.
European Union Reference Laboratory - Antimicrobial Resistance (EURL-AR) (2012): Protocol for PCR Amplifi cation of mecA, mecC (mecALGA251), spa and pvl Recommended by the EURL-AR, 2nd Version. EURL-AR/National Food Institute, pp. 1-5.
Fisher, E.L., Otto, M. & Cheung, G.Y.C. (2018): Basis of virulence in enterotoxin-mediated staphylococcal food poisoning. Front. Microbiol., 9, 436.
Hallin, M., Friedrich, A.W. & Struelens, M.J. (2009): spa typing for epidemiological surveillance of Staphylococcus aureus . -in: Caugant, D. (Ed.) Molecular epidemiology of microorganisms (Methods in Molecular Biology, vol. 551). Humana Press, Totowa, NJ, pp. 189-202.
Koreen, L., Ramaswamx S.V., Graviss, E.A., Naidich, S., Musser, J.M. &. Kreiswirth, B.N. (2004): spa typing method for discriminating among Staphylococcus aureus isolates: implications for use of a single marker to detect genetic micro- and macrovariation. J. Clin. Microbiol., 42, 792-799.
- Search Google Scholar
- Export Citation
)| false ( , Koreen, L. , Ramaswamx S.V. , Graviss, E.A. , Naidich, S. &. Musser, J.M. Kreiswirth, B.N. 2004): spa typing method for discriminating among. , Staphylococcus aureusisolates: implications for use of a single marker to detect genetic micro- and macrovariation 42, 792- 799.
Li, G., Wu, S., Luo, W., Su, Y., Luan, Y. & Wang, X. (2015): Staphylococcus aureus ST6-t701 isolates from foodpoisoning outbreaks (2006-2013) in Xi’an, China. Foodborne Pathog. Dis., 12, 203-206.
Manukumar, H.M. & Umesha, S. (2017): MALDI-TOF-MS based identification and molecular characterization of food associated methicillin-resistant Staphylococcus aureus. Sci. Rep., 7, 1-16.
Mossong, J., Decruyenaere, F., Moris, G., Ragimbeau, C., Olinger, C.M..&,Weicherding, P. (2015): Investigation of a staphylococcal food poisoning outbreak combining case-control, traditional typing and whole genome sequencing methods, Luxembourg, June 2014. EuroSurveillance, 20, 30059.
- Search Google Scholar
- Export Citation
)| false ( , Mossong, J. , Decruyenaere, F. , Moris, G. , Ragimbeau, C. .&, Olinger, C.M. Weicherding, P. 2015): Investigation of a staphylococcal food poisoning outbreak combining case-control, traditional typing and whole genome sequencing methods, Luxembourg, June 2014. , 20, 30059.
MSZ EN ISO 6888-1:2008: Élelmiszerek és takarmányok mikrobiológiája. Horizontális módszer a koágulazpozitív sztafilokokkuszok (Staphylococcus aureus es mas fajok) számának meghátározasára. 1. rész: Baird-Parker- agar taptalajos eljárás (ISO 6888-1:1999). Microbiology of food and animal feeding stuffs. Horizontal method for the enumeration of coagulase-positive staphylococci (Staphylococcus aureus and other species). Part 1: Technique using Baird-Parker agar medium (ISO 6888-1:1999).
Ortega, E., Abriouel, H., Lucas, R. & Gálvez, A. (2010): Multiple roles of Staphylococcus aureus enterotoxins: pathogenicity, superantigenic activity, and correlation to antibiotic resistance. Toxins, 8, 2117-2131.
Qi, C. & Sangma, X. (2019): Genotypes, enterotoxin gene profiles, and antimicrobial resistance of Staphylococcus aureus associated with foodborne outbreaks in Hangzhou, China. Toxins, 11(6), 307.