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Kit for detecting ocean incretion harass chemical substance, preparation method and application

A chemical substance and endocrine technology, applied in the direction of measuring devices, scientific instruments, analytical materials, etc., can solve the problems that fish are not distributed, can not be used to detect the pollution degree of chemical substances, etc., and achieve the effect of easy operation

Inactive Publication Date: 2009-04-01
OCEAN UNIV OF CHINA
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although foreign countries have also established enzyme-linked immunosorbent assay kits for marine fish vitellogenin, the fish detected by these kits are not distributed in my country, and cannot be used to detect the pollution degree of marine endocrine disrupting chemicals in my country

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0010] The following steps are involved in the preparation of the kit for the detection of marine endocrine disrupting chemicals:

[0011] (1) Preparation of pure sea bream vitellogenin

[0012] 10mg 17β-estradiol (17β-estradiol, E 2 ) was dissolved in 0.5mL of absolute ethanol, then 0.5mL of peanut oil was added, and mixed thoroughly to prepare 17β-estradiol emulsion. Red sea bream was injected intraperitoneally with 17β-estradiol emulsion, the injection dosage was 0.5mL / strip, blood was collected from the tail vein after 10 days, centrifuged at 8000g at 4°C for 5min, and vitellogenin-containing plasma was obtained. Take 1 mL of plasma and add it to a molecular sieve chromatography column, the filler is Sephacryl S-300, and it is eluted with 25mM Tris-HCl (pH7.5) containing 0.07M NaCl, and the sample containing red sea bream vitellogenin is collected for ion exchange chromatography. The filler is DEAE Sepharose Fast Flow, and 25mM Tris-HCl containing 0.1M and 0.2M NaCl is u...

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Abstract

The invention discloses a kit for detecting that the internal secretion of the sea disturbs chemical substances, and a preparation method and applications thereof. The preparation method comprises the following steps: when in preparation, firstly, the pure product of the vitellogenin of snapper is prepared; secondly, the polyclonal antibody of rabbit anti-vitellogenin of snapper is prepared; and thirdly, one bottle of pure vitellogenin of the snapper, one bottle of the polyclonal antibody of the rabbit anti-vitellogenin of the snapper, one block of blank 96-hole enzyme-linked immunosorbent assay board and respectively one bottle of the serum of a negative contrasting group, coating liquid, confining liquid, sample diluent, washing liquid, color development liquid, terminator and goat anti-mouse secondary antibody marked by horse radish peroxidase are filled into a box body together, thus obtaining the kit for detecting that the internal secretion of the sea disturbs the chemical substances. The kit for detecting that the internal secretion of the sea disturbs the chemical substances can be applied to the detection that the internal secretion disturbs the chemical substances and can quantitatively detect the vitellogenin in the blood of the snapper, liver tissues and the culture solution of hepatic cells, with sensitivity, accuracy and convenience.

Description

technical field [0001] The invention relates to a kit for detecting marine endocrine disrupting chemical substances, a preparation method and an application. Background technique [0002] In the last ten years, scientists have discovered that some pollutants entering aquatic ecosystems can disrupt the normal function of wildlife and human endocrine systems. This type of compound may be naturally occurring, such as soybean genistein, genistein, etc.; it may also be a synthetic compound, such as alkylphenols, polychlorinated biphenyls, and pesticides; Some are by-products of industrial production, such as dioxins, which are collectively referred to as endocrine disrupting chemicals. Endocrine disrupting chemicals can interfere with a series of biological processes such as the synthesis, release, transport, metabolism, binding of hormones and receptors, and expression of functions in humans and animals, thereby disrupting the endocrine system, nervous system and immune system ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543
Inventor 邴欣汝少国潘宗保
Owner OCEAN UNIV OF CHINA
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