Indirect competitive ELISA (Enzyme Linked Immunosorbent Assay) method based on multi-epitope tandem peptides and used for synchronously detecting staphylococcal enterotoxin A (SEA) and staphylococcal enterotoxin G (SEG)

Abstract

The invention belongs to the technical field of immunoassay and particularly relates to an indirect competitive ELISA (Enzyme Linked Immunosorbent Assay) method based on multi-epitope tandem peptides and used for synchronously detecting staphylococcal enterotoxin A (SEA) and staphylococcal enterotoxin G (SEG). The method mainly comprises preparation of SEA and SEG multi-epitope tandem peptide peptides (including antigens), preparation of anti-rabbit SEA and SEG antibodies, sample pretreatment and establishment of the indirect competitive ELISA detection method based on multi-epitope tandem peptides. The indirect competitive ELISA method comprises the following steps: (1) enveloping; (2) plate washing; (3) sealing; (4) plate washing; (5) adding standard substances, samples to be detected and specific antibodies in sequence; (6) plate washing; (7) adding ELIAS secondary antibodies; (8) plate washing; (9) developing and ending reaction. The method provided by the invention has the characteristics of high specificity, high sensitivity and good repeatability, is suitable for quick, synchronous and quantificational detection of SEA and SEG in foods such as milk and dairy products.

Description

Technical field [0001] The invention belongs to the technical field of immunoassays, and relates to a rapid, specific, sensitive and simultaneous detection method of Staphylococcus aureus type A and G enterotoxin in foods such as milk and dairy products, and specifically is an indirect competition based on multi-epitope tandem peptides ELISA method. Background technique [0002] Staphylococcus aureus (Staphylococcus aureus, referred to as Staphylococcus aureus) is an important food-borne pathogen, which is widely present in air, water, dust, and human and animal excrement. Staphylococcus aureus is easy to grow and reproduce and secrete enterotoxin (Staphylococcal Enterotoxin, SE) in foods with high water content, protein and starch, especially milk and dairy products. Since SE has strong thermal stability and toxicity, food contaminated with Staphylococcus aureus can be killed after heat treatment, but the SE produced by it is still pathogenic, and only a small amount of SE can ...

AI Technical Summary

Problems solved by technology

High-quality ELISA kits have been developed abroad for the detection of SE in foods such as milk and meat products, but they are expensive

In 2010, my country also designated the indirect ELISA method as the national standard method for detecting classic SE, but the detection range is limited, and only classic enterotoxins can be detected. There are blind spots in the detection of new enterotoxins, and the detection specificity and sensitivity are not satisfactory. false positive

At present, the relevant SE antigen detection methods declared in China can only detect classical enterotoxin, and the quality of detection reagents needs to be further improved

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