研究目的
To develop a rapid and sensitive MNP-based immunofluorescence method for staphylococcal enterotoxin A (SEA) determination in milk with low detection limit and without preliminary treatment of milk samples.
研究成果
The developed MNP-based immunofluorescence method is rapid, sensitive, and suitable for SEA detection in milk without preliminary sample treatment. It offers advantages such as short assay time, small sample volume, and effective magnetic separation.
研究不足
The method's sensitivity may be affected by the fat content in milk, with higher fat content leading to higher detection limits. The presence of nonspecific milk proteins could also influence the assay's performance.
1：Experimental Design and Method Selection
The study involved the development of a competitive immunoassay for SEA detection using magnetic nanoparticles (MNPs) modified with amino groups and conjugated with monoclonal anti-SEA antibody. The assay was optimized for MNP-Ab amount and fluorescent conjugate concentration.
2：Sample Selection and Data Sources
Milk samples spiked with SEA were used to test the immunoassay. The samples were prepared by diluting fresh milk with phosphate buffer.
3：List of Experimental Equipment and Materials
Magnetic nanoparticles, ATTO620NHS ester, monoclonal anti-SEA antibody, SEA, TEM for nanoparticle characterization, fluorescence spectrophotometer for conjugate analysis.
4：Experimental Procedures and Operational Workflow
MNPs were synthesized, modified with APTES, and conjugated with anti-SEA antibody. SEA was conjugated with ATTO620NHS. The competitive immunoassay was performed with optimized amounts of MNP-Ab and fluorescent conjugate, followed by magnetic separation and fluorescence measurement.
5：Data Analysis Methods
Fluorescence intensity was measured to determine the concentration of SEA in samples. The detection limit and linear range of the assay were calculated.
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