Volume 8, Issue 4 (10-2020)                   JoMMID 2020, 8(4): 132-136 | Back to browse issues page

XML Print

Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Golafrouz H, Ahari H, Anvar S A, Shahbazzadeh D. Detection of Staphylococcus aureus Enterotoxin A (SEA) Using Dot-ELISA in Milk Samples. JoMMID 2020; 8 (4) :132-136
URL: http://jommid.pasteur.ac.ir/article-1-232-en.html
Detection of Staphylococcus aureus Enterotoxin A (SEA) Using Dot-ELISA in Milk Samples
Haniyeh Golafrouz , Hamed Ahari * , Seyed Amirali Anvar , Delavar Shahbazzadeh
Department of Agricultural Sciences and Food Technology, Science and Research Branch, Islamic Azad University, Tehran, Iran
Abstract:   (2710 Views)
Introduction: Staphylococcus aureus enterotoxin A (SEA) is one of the most common causes of staphylococcal food poisoning. Due to the simplicity and no requirement for laboratory apparatuses, dot-ELISA is a choice method for detecting Staphylococcal enterotoxins. The present study aimed to develop a dot-ELISA for the detection of SEA. Methods: Nitrocellulose membranes were coated with the SEA antibody and blocked by the addition of 3% bovine serum albumin (BSA) blocking buffer. After 1 h incubation and washing the membranes, milk samples and the positive control (SEA, 50 ng/ml) were added to the membranes and incubated for 1 h. The membranes were then washed and incubated for 45 min with HRP-conjugated SEA, followed by the addition of TMB. Results: Our dot-ELISA could detect amounts of ≥ 50 ng/ml of SEA in the milk samples. Of the 30 raw milk samples randomly purchased from dairy product stores in District 3, Tehran, 5 (16%) contained SEA ≥ 50 ng/ml by the dot-ELISA. Conclusion: The dot-ELISA showed to be a reliable method for the preliminary screening of milk samples for SEA contamination. This method is cost-effective, fast, and does not require an ELISA-reader device.
Keywords: dot-ELISA, Staphylococcus aureus Enterotoxin A (SEA), Raw milk
Full-Text [PDF 810 kb]   (1355 Downloads)    
Type of Study: Original article | Subject: Diagnostic/screening methods and protocols
Received: 2020/01/17 | Accepted: 2020/10/19 | Published: 2021/02/13
References
1. LeChevallier MW, Seidler RJ. Staphylococcus aureus in rural drinking water. Appl Environ Microbiol. 1980; 39 (4): 739-42. [DOI:10.1128/AEM.39.4.739-742.1980]
2. Honeyman AL, Friedman H, Bendinelli M. Staphylococcus aureus Infection and disease. USA: Kluwer Academic Publisher, New York; 2001. [DOI:10.1007/b111097]
3. Le Loir Y, Baron F, Gautier M. Staphylococcus aureus and food poisoning. Genet Mol Res. 2003; 2 (1): 63-76.
4. Argudin MA, Mendoza MC, Rodicio MR. Food Poisoning and Staphylococcus aureus Enterotoxins. Toxins. 2010; 2 (7):1751-73. [DOI:10.3390/toxins2071751]
5. WHO estimates of the global burden of foodborne diseases, burden epidemiology reference group 2007-2015. https://www.who.int/foodsafety/publications/foodborne_disease/fergreport/en/.
6. Wu S, Duan N, Gu H, Hao L, Ye H, Gong W, Wang Z. A review of the methods for detection of Staphylococcus aureus enterotoxins. Toxins. 2016; 8 (7): 176-195. [DOI:10.3390/toxins8070176]
7. Thomas D, Chou S, Dauwalder O, Lina G. Diversity in Staphylococcus aureus enterotoxins. Chem Immunol Allergy. 2007; 93: 24-41. [DOI:10.1159/000100856]
8. Sospedra I, Marin R, Manes J, Soriano JM. Rapid whole protein quantification of staphylococcal enterotoxin B by liquid chromatography. Food Chem. 2012; 133 (1): 163-6. [DOI:10.1016/j.foodchem.2011.12.083]
9. Salmain M, Ghasemia M, Boujday S, Spadavecchia J, Te'cher C, Val F, Le Moignee V, Gautier M, Briandet R, Pradier CM. Piezoelectric immunosensor for direct and rapid detection of staphylococcal enterotoxin A (SEA) at the ng level. Biosens Bioelectron. 2011; 29 (1): 140-4. [DOI:10.1016/j.bios.2011.08.007]
10. Johler S, Weder D, Bridy C, Huguenin MC, Robert L, Hummerjohann J, et al. Outbreak of staphylococcal food poisoning among children and staff at a Swiss boarding school due to soft cheese made from raw milk. J Dairy Sci. 2015; 98 (5): 2944-8. [DOI:10.3168/jds.2014-9123]
11. Gadyari F, Sattari M, Boroumand MA, Yaghoubi R, Sepehriseresht S, Purgholi L. Molecular tracing of A, B, C, and D enterotoxins of Staphylococcus aureus in clinical strains separated from burn patients of Motahhari Hospital in Tehran. Iran J Medical Microbiol. 2011; 5 (5): 20-7. (in Persian)
12. Sharif AS, Sattari M, Moradi M, Shahrokhabad R. Detection of Staphylococcus aureus enterotoxin genes A and B in clinical samples of the patients referring to the medical centers of Kerman and Rafsanjan cities by PCR Technique. J Rafsanjan Uni Med Sci. 2012; 11: 128-36. (in Persian)
13. Ahmadi M, Rohani SMR, Ayremlou N. Detection of Staphylococcus aureus in milk by PCR. Comp Clin Path. 2010; 19 (1): 91-4. [DOI:10.1007/s00580-009-0901-0]
14. Abbassi S, Taei S, Zamanzad B. The prevalence of methicillin-resistant Staph. aureus strains producing enterotoxin A and B. Tehran Univ Med J. 2016: 73(11); 778-83. (in Persian)
15. Barati B, Saadati M, Bahmani MK. Isolation and Detection of Enterotoxigenic Staphylococcus aureus Type A by Multiplex PCR. J Mil Med. 2006; 8 (2): 119-28.
16. Kuang H, Wang W, Xu L, Ma W, Liu L, Wang L, et al. Monoclonal antibody-based Sandwich ELISA for the detection of Staphylococcal enterotoxin A. Inter J Environ Res Public Health. 2013; 10 (4): 1598-608. [DOI:10.3390/ijerph10041598]
17. Nouri A, Ahari H, Shahbazzadeh D. Designing a direct ELISA kit for the detection of Staphylococcus aureus enterotoxin A in raw milk samples. Int J Biol Macromol. 2018; 107 (Pt B): 1732-7. [DOI:10.1016/j.ijbiomac.2017.10.052]
18. Soejima T, Nagao E, Yano Y, Yamagata H, Kagi H, Shinagawa K. Risk evaluation for staphylococcal food poisoning in processed milk produced with skim milk powder. Inter J Food Microbiol. 2007; 115 (1): 29-34. [DOI:10.1016/j.ijfoodmicro.2006.10.009]
19. Fujikawa H, Morozumi S. Modeling Staphylococcus aureus growth and enterotoxin production in milk. Food Microbiol. 2006; 23 (3): 260-7. [DOI:10.1016/j.fm.2005.04.005]
20. Madahi H, Rostami F, Rahimi E, Safarpoor S. Prevalence of enterotoxigenic Staphylococcus aureus isolated from chicken nugget in Iran. Jundishapur J Microbiol 2014; 7 (8): 10237. [DOI:10.5812/jjm.10237]
21. Singh M, Agrawal RK, Singh BR, Mendiratta SK, Agarwal RK, Singh MK, et al. Development and evaluation of simple Dot-Blot assays for rapid detection of Staphylococcal enterotoxin-A in food. Indian J Microbiol. 2017; 57 (4): 507-11. [DOI:10.1007/s12088-017-0671-3]
Add your comments about this article : Your username or Email:
CAPTCHA
Send email to the article author

Rights and permissions
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.