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Preparation method of kit for detecting fish livetin source of carp family

A vitellogenin and kit technology, applied in biological testing, measuring devices, scientific instruments, etc., can solve problems such as few or no fish, unrepresentative, and expensive kits

Inactive Publication Date: 2003-02-05
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

Method used

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Embodiment Construction

[0008] The steps are as follows:

[0009] (1) Induction and separation and purification of vitellogenin standard samples

[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 (pH7.5) buffer, then 2 mL of serum sample was added, and gradient elution was carried out with Tris-HCl (pH7.5) buffer containing NaCl, and a partial collector was used to collect the eluted Remove the liquid, and determine whether each tube contains vitellogenin...

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Abstract

The invention discloses the method for preparing the kit for detecting vitellogenin of cyprinid. 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. With a period of time of induction, the blood serum of the fish blood is obtained through the centrifugal procedure. 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 vitellogenn. The weighted freeze-dried power is taken to dissolve in the double steamed water with Freund's adjuvant being added to emulsify. The polyclonal antibody is prepared through the immune giant blanc. The invention can be used for quantificationally detecting vitallogenin in blood, liver and body of cyprinid with accuracy.

Description

technical field [0001] The present invention relates to research on ecological safety and risk assessment of estrogens, in particular to a method for quantitatively detecting the content of vitellogenin (VTG) in the blood, liver tissue and fish body of carps based on the principle of enzyme-linked immunosorbent adsorption The preparation method of the kit. Background technique [0002] At present, humans use a large number of pesticides and herbicides to increase agricultural production, and the application of many chemical products has also significantly improved people's living standards. But the extensive use of these artificial chemicals has also caused great harm to our environment and human health itself. In recent years, more and more studies have shown that the fertility of many wild animals has declined, wild communities have degraded, the number and quality of human sperm have decreased, and the incidence of breast cancer and testicular can...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/531G01N33/535G01N33/558G01N33/574
Inventor 徐盈梁勇
Owner INST OF AQUATIC LIFE ACAD SINICA
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