研究目的
Developing a rapid, sensitive, and label-free method for the determination of aflatoxin M1 (AFM1) levels in milk samples using White Light Reflectance Spectroscopy (WLRS) immunosensor.
研究成果
The WLRS biosensor developed for the determination of AFM1 in milk samples is characterized by high detection sensitivity, short analysis time, and low cost of consumables. It could detect AFM1 in whole milk at concentrations as low as 6 pg/mL, well below the maximum allowable limit set by EU and other regulatory authorities. The sensor is suitable for rapid and sensitive detection of AFM1 at the point-of-need.
研究不足
The presence of milk, even 100-times diluted, had a significant effect on the signal recorded in real-time due to light absorption or scattering from milk. However, by introducing a washing step, it was possible to monitor the specific reactions.
1:Experimental Design and Method Selection:
A competitive immunoassay format was followed, including functionalization of the chip through immobilization of an AFM1-protein conjugate. The assay involved running mixtures of anti-AFM1 specific antibody with calibrators or samples over the biochip, followed by reaction with biotinylated secondary antibody and streptavidin.
2:Sample Selection and Data Sources:
Milk samples from different origins (cow, goat, sheep, and human) were used without any pretreatment.
3:List of Experimental Equipment and Materials:
The WLRS sensing system consists of a USB powered & controlled measurement set-up (including reflection probe, light source, spectrometer) and a biochip (Si chip with a SiO2 layer).
4:Experimental Procedures and Operational Workflow:
The assay cycle included equilibration with assay buffer, primary immunoreaction, washing, reaction with biotinylated secondary antibody and streptavidin, and regeneration.
5:Data Analysis Methods:
The reflected spectrum was recorded continuously and processed by dedicated software to transform the spectral shift to effective biomolecular adlayer thickness increase.
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