Research On Immunochip Techniques For The Detection Of Fumonision B₁ And Stapylociccus Enterotoxin A,B

Immunochip is a kind of protein-chip. It is a microassay technique that involves coating protein on glass slide. There are some advantages than other traditional techniques as a tool for detecting lots of biotoxin. So It is becoming popular attention. Immunochips have been developed for the detection of the fumonision B1 and stapylociccus enterotoxin A,B in the paper.The first step in developing immunochips is to coat the surface of carriers with antibody or antigen by using the glass sliders as carriers, which are treated with glutaraldehyde to provide aladehyde. The study accomplishes the immobilization of antibody or antigen on the carriers through the combination of aldehyde and the free amido of antibody or antigen.The antibody combining method was adopted in the process of detecting fumonision B₁. Before detection, the samples and Cy3 conjugated fumonision B₁-BSA were simultaneously added to the immunochip. The fumonision B₁ contained in samples and the Cy3 conjugated fumonision B₁-BSA will competitivily combine the antibody on carriers. As the conjugated fumonision B₁ its quantity is detected. If it is higher concertration of fumonision B₁ in samples, the quantity of Cy3 conjugated fumonision B₁-BSA will decline, then we will get low fluorescent intensity. It is the method that we use to complete the detection. The detection limits of fumonision B₁ in our study is 1μg/ml.In the process of detecting stapylociccus enterotoxin A,B, the antibody sandwich method was adopted. And the conditions of immunochips which influence the detection results were optimized. Before detection , stapylociccus enterotoxin A,B were added to the chips. The uncombined stapylociccus enterotoxins were washed away, then the fluorescence conjugated antibodies were added to form the sandwich structure. The more stapylociccus enterotoxins in samples, the more combined antibody. As a result a strong fluorescent intensity can be measured of stapylociccus enterotoxin A,B. The results showed that the fluorescence intensity increased with the increasing concentration of samples. When the concentrations of SEs were too high, the fluorescence intensity did not increase any more. And we could detect stapylociccus enterotoxin A,B on the same glass slide at the same time. A series of optimizations of testing conditions was carried out for the detection of stapylociccus enterotoxin A,B. The detection limits of SEA, SEB were all 1. 001μg/ml respectively.The paper indicates that with the experiment moving on , the progress and maturity of protein-chip techniques, Immunochip technique is becoming one of the important and popular methods in the detection of food safety fleetness. It is predicted that this high-tech will be used in many more fields.