Test kit for qualitatively detecting fish vitellogenin by utilizing lipovitellin antibody

Abstract

The invention relates to a test kit for qualitatively detecting fish vitellogenin by utilizing lipovitellin antibody. The test kit comprises pure product of goldfish lipovitellin and anti-rat goldfish lipovitellin polyclonal antibody. The preparation method of the test kit comprises the following steps: obtaining goldfish lipovitellin through purifying at first; then preparing anti-rat goldfish lipovitellin polyclonal antiserum through immune rats; and further purifying anti-rat goldfish lipovitellin polyclonal antibody. According to the invention, the test kit can be used for screening environmental estrogen materials and monitoring the situation of water environment which is polluted by the environmental estrogen materials, so that vitellogenin in fish blood, overall uniform serous fluid, skin mucus, liver tissues and hepatic cell nutrient solution can be monitored sensitively and conveniently. The test kit can be used for qualitatively detecting cyprinidae fish vitellogenin, and has the same monitoring sensitivity to the fish vitellogenin of a far provenance relationship fish. The biggest advantage of the test kit is that the test kit has a very wide application scope, and the minimum monitoring limit can reach 100ng / ml.

Description

technical field [0001] The invention belongs to the field of ecological detection, and in particular relates to a kit for qualitatively detecting fish vitellogenin by using a vitellin phosphoprotein antibody, and a preparation method and application thereof. Background technique [0002] In recent years, the effects of environmental estrogens on the reproductive system of wild animals and humans has received extensive attention. In order to reduce the harm of environmental estrogen to organisms and humans, it has become a current research hotspot to establish a fast and effective screening method to screen out chemical substances with environmental estrogen effects from environmental pollutants. The United States, the European Union and Japan have successively established environmental estrogen screening and evaluation systems using fish as model organisms. Vitellogenin (Vtg), as an important biological screening index, has been widely used. [0003] Vitellogenin is the pre...

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Problems solved by technology

[0005] At present, a variety of enzyme-linked immunosorbent assay kits and radioimmunoassay kits for fish vitellogenin have been established at home and abroad, but these kits have high requirements for instruments, and there are many influencing factors in the experimental process; Among them, the immunological and structural properties of vitellogenin are quite different. Even the Vtg of fish with similar species relationship has strong heterogeneity. Usually, vitellogenin antibody can only be used for vitellogenin of specific fish species. However, there is no or only very weak cross-reactivity with other fish species, which means that it is necessary to develop species-specific antibodies; and due to the large differences in the geographical distribution of fish, it is necessary to prepare and use easily and require less equipment. Not high, able to overcome interspecies differences, a detection kit for fish vitellogenin with a wide range of applications

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