Immunochemical assay with monoclonal antibodies for detection of staphylococcal enterotoxin H

Staphylococcal enterotoxins cause food poisoning of various degrees of severity. For milk and meat products, there is a high probability of contamination with staphylococcal enterotoxin H (SEH). In this regard specific and sensitive methods are required to be developed for its detection and monitori...

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Bibliographic Details
Published in:Yàowu shi͡p︡in fenxi 2018-04, Vol.26 (2), p.741-750
Main Authors: Rudenko, Natalia Vasilyevna, Karatovskaya, Anna Petrovna, Noskov, Anatolyi Nikolaevich, Shepelyakovskaya, Anna Olegovna, Shchannikova, Margarita Pavlovna, Loskutova, Irina Vladimirovna, Artyemieva, Olga Anatolievna, Nikanova, Daria Alexandrovna, Gladyr, Elena Alexandrovna, Brovko, Fedor Alexandrovich
Format: Article
Language:English
Subjects:
Cow's milk
Cross-reactivity
Enzyme immunoassay
Food
Food contamination
Food contamination & poisoning
Food poisoning
Food production
Food products
Genes
Immunoassay
Immunoblotting
Mastitis
Meat
Meat products
Milk
Monoclonal antibodies
Original
Polyclonal antibodies
Proteins
Sandwich enzyme immunoassay
Staphylococcal enterotoxin H
Staphylococcal enterotoxins
Staphylococcus aureus
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Summary:Staphylococcal enterotoxins cause food poisoning of various degrees of severity. For milk and meat products, there is a high probability of contamination with staphylococcal enterotoxin H (SEH). In this regard specific and sensitive methods are required to be developed for its detection and monitoring. In this work, the gene seh was expressed and a preparation of recombinant toxin was obtained. Using hybridoma technology, a panel of high-affinity monoclonal antibodies (mAbs) to SEH was produced. The antibodies were characterized and shown to have no cross-reactivity towards the main staphylococcal enterotoxins (A, B, C1, D, E, G and I). Based on these mAbs, a method for specific and quantitative detection of SEH was developed in the format of sandwich enzyme immunoassay (linear range, 0.2–3 ng/ml). All the mAbs produced revealed SEH by immunoblotting. Immunochemical analysis of the culture fluids of staphylococcal isolates obtained from the milk of mastitis-infected cows by immunoblotting and sandwich enzyme immunoassay demonstrated the conformity of these methods. Using the developed method, the toxin was revealed in blood serum and liquid food products practically to 100%. From non-liquid foods, it was shown to be extracted to a maximum with a buffer of pH 4.0–4.5. [Display omitted] •18 hybridomas, secreting high-affinity monoclonal antibodies to SEH, were obtained.•A sandwich immunoassay was developed for the quantitation of SEH.•The detection ranges of SEH were 0.2–3.0 ng/ml.•Measured SEH, secreted by S. aureus isolates, obtained from mastitis-infected cows.•The developed method effectively detects SEH in food and blood serum.
ISSN:1021-9498
2224-6614
DOI:10.1016/j.jfda.2017.10.011