Preparation method of reagent kit for quickly detecting estrogenoid toxicity

A detection kit and estrogen technology are applied in the field of preparation of estrogen-like toxicity rapid detection kits, which can solve the problems of unrepresentativeness, expensive kits, and difficulty in promotion.

Inactive Publication Date: 2004-10-06
INST OF AQUATIC LIFE ACAD SINICA
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Problems solved by technology

[0004] At present, several kinds of enzyme-linked immunosorbent assay kits and radioimmunoassay kits for vitellogenin of teleosts have been established abroad, but the distribution of fish that can be detected by the kits is few or none in China, which is not representative , and the price of the kit is extremely expensive, it is difficult to promote

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Embodiment Construction

[0008] The steps are as follows:

[0009] (1) Induction and separation and purification of the vitellogenin standard sample used in the present invention

[0010] Preparation of 17α-ethinylestradiol (17α-ethinylestradiol, EE 2 ) solution: 100 mg of 17α-ethynyl estradiol was first dissolved in 5 mL of acetone, then dissolved in 10 mL of soybean oil, and the acetone was blown dry with nitrogen. Intraperitoneal injection into the fish body, the dose is 5mg / kg, after 5-7 days of exposure, the blood of the fish is taken, and the serum is obtained by centrifugation at 4°C. The serum was passed through an ion-exchange chromatography column, and the filler was DEAE Sepharose CL-6B. The chromatographic column was first equilibrated with 0.025mol / L Tris-HCl (pH 7.5) buffer solution, then 2mL serum sample was added, gradient elution was carried out with Tris-HCl containing NaCl, (pH 7.5) buffer solution, and a partial collector was used to collect Eluate, and electrophoresis to determ...

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Abstract

The invention discloses the method for preparing the kit for fast detecting toxicity of generic estrogen. The first step is to induce, separate and purify the vitellogenin. The prepared solution of lynoral is injected into the stomach cavity of the cyprinid. Then, the freeze-dried powder is prepared by using the separated purification of ion exchange column and desalting of dialysis. The next step is to prepare polyclonal antibody of vitellogenin by using the dissolved freeze-dried powder with Freund's adjuvant being added to emulsify and through the immune giant blanc. The final step is to prepare polyclonal antibody of vitellogenin with the marker of biotin. The invention can be used for quantificationally detecting vitellogenin in blood, liver and body of cyprinid with accuracy.

Description

technical field [0001] The present invention relates to the research on the ecological safety and risk assessment of estrogens, in particular to a biotin-avidin enzyme-linked immunosorbent adsorption principle for the quantitative detection of carps blood, fish liver tissue, fish liver cells A method for preparing a kit for rapid detection of estrogen-like toxicity in culture medium and fish body vitellogenin (VTG) content. Background technique [0002] Endocrine disruptors are those exogenous compounds that can interfere with the production, secretion, transport, metabolism and action of endogenous hormones that maintain endocrine balance in the body and regulate growth and development. They can interfere with the function of the endocrine system of organisms, thereby affecting the growth, development and reproduction of organisms. Humans synthesize many new compounds every year, and while they bring convenience to people's lives, they also cause...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/53G01N33/531G01N33/535G01N33/558G01N33/74
Inventor 徐盈梁勇
Owner INST OF AQUATIC LIFE ACAD SINICA
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