Preparation and detection methods of kit for detecting chub vitellogenin (VTG)

Abstract

The invention relates to preparation and detection methods of a kit for detecting chub vitellogenin (VTG). The preparation method comprises the following steps of: inducing the chub VTG, and separating and preparing purified chub VTG freeze-dried powder; preparing a specific anti-VTG polyclonal antibody, namely a primary antibody; and placing the purified chub VTG freeze-dried powder, the primary antibody, an enzyme-labeled secondary antibody and an ELISA (enzyme linked immunosorbent assay) plate into the kit to finally obtain the kit for detecting the chub VTG. The detection method comprises the following steps of: using the purified chub VTG freeze-dried powder as a standard protein, and adding a diluted sample solution into the ELISA plate; adding the primary antibody into the ELISA plate for heat insulation reaction; adding the enzyme-labeled secondary antibody into the ELISA plate for heat insulation reaction; and adding a 3,3,5,5-tetramethyl benzidine substrate solution for color development reaction, determining an absorbance value by an ELISA reader, and calculating the concentration of the VTG in a chub sample. According to the detection method disclosed by the invention, the content of the VTG in chub serum, homogenate and the like can be quickly tested, the analysis sensitivity is high, the test time is short, and the detection method further has the advantages of time saving and labor saving and is suitable for analyzing and screening a large number of samples.

Description

Technical field [0001] The invention belongs to the field of freshwater product environmental analysis, and in particular relates to a preparation and detection method of a kit for detecting silver carp vitellogenin. Background technique [0002] Estrogens are a class of environmental chemicals that have estrogenic activity and can mimic the function of estrogen. Studies have shown that many human reproductive disorders, developmental abnormalities and certain cancers are related to estrogen-like hormones. The ultimate fate of these pollutants in the environment is various water bodies, and they are enriched in fish through the food chain. Vitellogenin (VTG) is a specific biomarker of estrogen-like pollution in fish. At present, the quantitative analysis of proteins mainly includes high pressure liquid chromatography (HPLC), capillary electrophoresis (CE) and liquid chromatography-mass spectrometry (LC-MS) and other chemical analysis methods. However, chemical analysis requires ...

AI Technical Summary

Problems solved by technology

At present, there are mainly chemical analysis methods such as high-pressure liquid chromatography (HPLC), capillary electrophoresis (CE) and liquid chromatography-mass spectrometry (LC-MS) for quantitative analysis of proteins. Concentration and other steps, complicated pre-treatment, time-consuming, laborious, low sensitivity, not suitable for analysis and screening of a large number of samples

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