Journal
FOOD CHEMISTRY
Volume 353, Issue -, Pages -Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.foodchem.2021.129481
Keywords
Staphylococcus aureus; Food-borne pathogen; Nanobody; Sandwich ELISA; Immuno-assay
Funding
- National Natural Science Foundation of China [31901779]
- Fundamental Research Funds for the Central Universities, Nankai University
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In this study, specific nanobodies recognizing Staphylococcus aureus were selected and an immuno-assay was established to detect S. aureus contaminations in foods, with a detection limit of 10 CFU/mL in milk samples. This research provides a basis for an easy, reproducible and effective immuno-assay for screening S. aureus contaminations in foods.
The interaction between conventional immunoglobulins (Igs) and the Ig-binding surface proteins of Staphylococcus aureus (S. aureus) have obstructed the development of immuno-assays to detect these bacteria. The current study aimed to select nanobodies (Nbs) recognizing specifically S. aureus and to establish an immuno-assay to uncover S. aureus contaminations in foods. An alpaca was immunized with an inactivated S. aureus strain followed by the construction of a Nb library from which four target-specific Nbs were retrieved. Subsequently, a sandwich ELISA employing the Nb147 and biotinylated-Nb147 pair to capture and to detect S. aureus, respectively, was established to possess a detection limit of 1.4 x 105 colony forming units (CFU)/mL. The dedicated immuno-assay has been verified by detecting 10 CFU/mL of S. aureus in milk samples after an 8 h-enrichment step. This study provides the basis of an easy, reproducible and effective immuno-assay to screen for S. aureus contaminations in foods.
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