Designing Dot ELISA kit for detection of Staphylococcus aureus SEA enterotoxin and Sandwich ELISA compare, its validation in pastry cream bread

Subject Areas : Journal of Comparative Pathobiology

, SA. Bathaei ¹ , , H. Ahari ² , , AA Anvar ^{3 *}

1 - .. M.c.s, Agricultural Sciences and Food Technology, Science and Research Branch, Islamic Azad University, Tehran, Iran
2 - Associate Professor, Sciences and Food Technology, Science and Research Branch, Islamic Azad University, Tehran, Iran
3 - Department of food hygiene, Science and Research Branch, Islamic Azad University, Tehran, Iran

Received: 2021-05-02 Accepted : 2021-05-02 Published : 2020-06-21

Keywords: Enterotoxin, Staphylococcus aureus, Dot ELISA, Kit,

Abstract :

In this study, detection and recognition of Staphylococcus aureus enterotoxin was performed by designing a Dot ELISA sensitive kit. In the present study, nitrocellulose membrane (NCM) with 0.22 micron pores was applied as the bed, and 1:500 diluted specific antibody of Staphylococcus aureus enterotoxin was coated on the NCM. Next, The HRP-tagged antibody was added with dilution of 1/2000, and the substrate was added gradually and rewashed. In this experiment, the samples were manually contaminated and then compared with the ELISA sandwich method as a gold standard. The kit is capable of detecting 50 ng/ml and higher of toxin A, and if the amount of toxin is less, it will report it negatively. In this test, of which 40 samples were collected and 4 positive cases were reported, it was revealed that about 10% of the collected samples were contaminated. Nowadays, using the Dot ELISA kit as a quick method with relative sensitivity and properties, it is possible to improve the repeatability of detecting Staphylococcus enterotoxin A in creamy candies.

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