Method for detecting soybean antigen beta-conglycinin

Abstract

The invention discloses a method for detecting soybean antigen beta-conglycinin. A direct ELISA method for detecting soybean antigen beta-conglycinin is established. The envelope antigen is determined to be 1mu g / mL by virtue of a square matrix experiment, the optimal dilution degree of the antibody is 1:400, the optimal confinement time is determined to be 58.31 minutes by virtue of the orthogonal response surface experiment design, the optimal reaction time of an enzyme-labeled antibody is 139.16 minutes, the optimal reaction time of substrate solution action is 19.43 minutes, the sensitivity is 1ug / ml, and the linear detection range is between 20ng / ml and 640ng / ml. The mean number of the variation coefficient (CV%) between the plates is 8.16 percent, the mean number of the variation coefficient (CV%) in the plates is 6.91 percent, and the cross reaction is smaller than 20 percent. The detection method disclosed by the invention has the characteristics of relatively simple sample pretreatment, rapidness, low cost, low instrumentation degree, short time consumption and the like, can be applied for detecting the content of glycinin in soybean and deep processing products and is applied to field monitoring and large-scale sample screening.

Description

technical field [0001] The invention belongs to the field of food and feed safety detection, and in particular relates to a detection method of soybean antigen β-conglycinin. Background technique [0002] Soybean is rich in protein and balanced amino acids, and is a high-quality plant-based protein source for humans and livestock. However, the phenomenon that the antigenic protein contained therein causes allergic reactions in humans and animals has attracted more and more attention. [0003] Soybean antigens refer to a class of proteins in soybeans that can cause allergic reactions in humans and young animals. Among them, β-conglycinin is one of the main soybean antigenic proteins, which hinders the wide application of soybean antigenic eggs in the food and feed industries. In livestock and poultry production, the intake of soybean-containing diets can cause allergic reactions such as diarrhea, growth retardation, and intestinal mucosal cell proliferation in young animals ...

AI Technical Summary

Problems solved by technology

High-performance liquid chromatography is time-consuming and the instrument is relatively expensive, so it is not suitable for actual production applications; the detection method of western blot has certain accuracy, but it takes a long time during the test, and The cost of the test is relatively high, so it is limited in some routine production testing; PCR technology (Polymerase Chain Reaction) is a molecular biology technique used to amplify specific DNA fragments, but in the later stage it is used in the analysis of DNA content and quality. PCR technology will be affected during the technical process such as extraction and extraction amount. During the PCR detection process, any factors that affect DNA content, quality, and extraction amount will affect the use of PCR technology, thus hindering the development and promotion of this technology.

[0005] The above methods all have certain efficiency and sensitivity, but are expensive and time-consuming, and are not suitable for rapid analysis of a large number of samples in production.

Enzyme-linked immunosorbent assay, which can be used for qualitative or quantitative analysis, has been widely recognized for its advantages of simple operation, rapidity, and suitability for batch detection. At present, the common detection methods include double-antibody sandwich method and competition method, but these methods require more There are many protein standards and their antibodies, and the detection cost is high. Among them, the indirect competitive sandwich ELISA method is not suitable for mass detection in the food and feed industry due to the cumbersome steps and waste of antigen reagents.