Enzyme linked immunosorbent assay kit for detecting progesterone and detection method thereof

An enzyme-linked immunosorbent reagent and detection method technology, which is applied in the field of enzyme-linked immunosorbent immunoassay kits for detecting progesterone in chicken tissue, can solve the problem of no progesterone in chicken, and achieve the effect of simple operation and sensitive detection

Inactive Publication Date: 2015-05-27
ZHENJIANG XIANCHUANG BIOTECH CO LTD
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AI Technical Summary

Problems solved by technology

However, there is no relevant report on the detection of progesterone residues in chicken meat in China. Therefore, it is necessary to establish a detection technology for progesterone residues in chicken tissues.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0027] Preparation of progesterone protein conjugates:

[0028] The carboxylated progesterone hapten derivative was obtained by the succinic anhydride method, and then 0.05 mmol was mixed with the carrier protein BSA at a binding ratio of 10:1 in 0.05 mol / L carbonate buffer solution (CBS) at pH 9.6, and then Add 0.15 mmol carbodiimide, stir and react at room temperature for 24 h, and finally dialyze in 0.2 mol / L PBS buffer solution with pH 7.6 for two days to remove unreacted hapten, and put the obtained protein conjugate solution at -20 Store at ℃ for later use.

[0029] Preparation of progesterone antibody:

[0030] Select healthy adult purebred BALA / C mice, take 50 μg of immune antigen prepared by coupling with protein and mix with the same amount of complete Freund's adjuvant, and use intraperitoneal injection for initial immunization, and then use the same dose of immune antigen plus the same amount every 3 weeks Incomplete Freund's adjuvant was used for the second and ...

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Abstract

The invention provides an enzyme linked immunosorbent assay (ELISA) kit for detecting progesterone and a detection method thereof. The kit is sensitive, accurate, quick in detection, is simple in operation, is strong in specificity and is suitable for detection of samples in large amount. The kit includes: an ELISA plate coated by a progesterone antigen, a progesterone standard sample, a progesterone antibody operating fluid, a progesterone enzyme label second antibody operating fluid, a substrate liquid A, a substrate liquid B, a stop buffer liquid, a concentrated diluent and a concentrated washing liquid. The principle of the ELISA kit for detecting progesterone is solid-phase indirect competitive ELISA. In the detection method, an extracted sample, the progesterone enzyme label second antibody operating fluid and the progesterone antibody operating fluid are added to corresponding microholes in the ELISA plate. After incubation for a certain time, the ELISA plate is washed and is added with the substrate liquid A and the substrate liquid B, and then a color developing agent develops a blue color under effect of enzymes. After the stop buffer liquid added, the color is turned into yellow from blue. An inversely proportional relationship is formed between the depth of the developed color and the content of the progesterone in the standard sample or the sample. The method can be directly used for detecting residual amount of the progesterone in chicken tissue.

Description

technical field [0001] The invention relates to the technical field of veterinary drug residue detection, in particular to an ELISA kit for detecting progesterone in chicken tissue. Background technique [0002] Progesterone, also known as progesterone, progesterone, progesterone, luteinizing hormone, progestin, progestin, or progesterone, is a hormone involved in the female menstrual cycle, pregnancy, and embryonic effects in humans and other animals. of steroids. Progesterone is a female hormone involved in the female menstrual cycle in humans and other animals, supporting pregnancy and embryo formation. Progesterone belongs to a class of hormones called progestins and is the most important progestin in humans. Hormone drugs can improve the feed conversion rate of animal food, and then achieve the purpose of weight gain, but the residual progesterone in the body will cause a series of problems, such as changes in sexual characteristics, premature puberty, etc., and the e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N21/31
CPCG01N33/743
Inventor 洪霞杜霞
Owner ZHENJIANG XIANCHUANG BIOTECH CO LTD
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