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A "one to two" novel sandwich immunoassay based on nanobodies for detection of staphylococcal enterotoxin A in food samples.
- Source :
-
Food Control . Jun2024, Vol. 160, pN.PAG-N.PAG. 1p. - Publication Year :
- 2024
-
Abstract
- Staphylococcal enterotoxin A (SEA), a virulence factor produced by Staphylococcus aureus (S. aureus), poses a serious threat to human health. The rapid and sensitive detection of SEA is crucial for preventing the spread of contaminated food and safeguarding public health. The conventional enzyme-linked immunosorbent assay (ELISA) has encountered limitations in detecting SEA due to the false-positives caused by the combination of the Fc-terminus of the antibodies and staphylococcal protein A (SpA) and its insufficient sensitivity that fails to meet current detection requirements. Therefore, nanobodies (Nbs) without Fc-terminus can replace traditional antibodies to prevent false-positives. We developed a "one to two" sandwich ELISA based on identical capture Nbs and double detection phage-displayed Nbs (OtTNb ELISA), which achieved detection with an LOD of 0.43 ng/mL in a linear range of 0.5–512 ng/mL. The sensitivity was over 3.4 times higher than that of the sandwich ELISA based on mAbs, and the linear range exceeded over 8 times. The OtTNb ELISA was successfully applied to the cross-reactivity test and the actual recovery of the sample. This study provides a sensitive tool for detecting staphylococcal enterotoxins (SEs) in food and offers insights into developing detection methods for various foodborne pathogens and toxins. [Display omitted] • The first design of an"One to Two" sandwich ELISA based on identical capture Nbs and double detection phage-displayed Nbs. • Drastically increase the load of HRP and the antigen-antibody binding sites. • The sensitivity of this assay was over 3.4 times higher than that of the sandwich ELISA based on mAbs. • This assay was achieved detection of SEA with an LOD of 0.43 ng/mL and a linear range of 0.5–512 ng/mL. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 09567135
- Volume :
- 160
- Database :
- Academic Search Index
- Journal :
- Food Control
- Publication Type :
- Academic Journal
- Accession number :
- 175773795
- Full Text :
- https://doi.org/10.1016/j.foodcont.2024.110313