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Detection kit of cow milk progesterone content

A technology for detecting kits and milk progesterone, which is applied in biological tests, measuring devices, material inspection products, etc., can solve the problems of high import costs and other problems, and achieve the effects of high repeatability, fast and accurate detection, and good specificity.

Inactive Publication Date: 2011-01-19
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no commercial quality-guaranteed milk-progesterone test kit available in the market. To promote milk-progesterone test kits in China, the current best way is to import enzyme-immune milk-progesterone test kits from abroad, but the import cost is expensive.

Method used

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  • Detection kit of cow milk progesterone content
  • Detection kit of cow milk progesterone content
  • Detection kit of cow milk progesterone content

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The preparation of embodiment 1 progesterone complete antigen

[0034] The succinic anhydride method was used to activate 11α-hydroxyprogesterone, and the carbodiimide method was further used to select BSA and KLH as immune antigen carriers and OVA as detection antigen carriers for coupling reaction. Further immunize animals with immune antigens to prepare antiserum, determine the optimal coating concentration of the detection antigen P-OVA, and indirect ELISA to detect whether the antiserum is specific to the progesterone residue in the complete antigen; Residues of the antibody, whether it can specifically bind to the free progesterone molecule itself. Comparing the immune effects of BSA and KLH two different proteins as immune antigen carriers, and selecting the best immune antigen, lays the foundation for the next step of preparing progesterone monoclonal antibody.

[0035] 1 Materials and methods

[0036] 1.1 Materials

[0037] Male BabL / C mice aged 6-8 weeks we...

Embodiment 2

[0081] Example 2 Preparation and preliminary identification of progesterone monoclonal antibody

[0082] Using the prepared P-BSA11α-hydroxyprogesterone complete antigen as the immune antigen to immunize mice, adopting chemical fusion method, using polyethylene glycol as the fusion agent, through the steps of fusion, screening, cloning, expanded culture, purification and identification, etc., to obtain Monoclonal antibodies against progesterone. Preliminary identification of parameters such as specificity, cross-reactivity, and relative affinity was carried out, and the best monoclonal antibody was selected from them for the establishment of ELISA detection method.

[0083] 1 Materials and methods

[0084] 1.1 Materials

[0085] Male BabL / C mice aged 6-8 weeks were purchased from Changchun Institute of Biological Products. SP2 / 0 cells. P-BSA is the complete antigen prepared in Example 1; horseradish peroxidase-labeled goat anti-mouse IgG and horseradish peroxidase-labeled ...

Embodiment 3

[0161] The preparation of embodiment 3 progesterone ELISA detection kit

[0162] 1. Kit composition and preparation

[0163] ① ELISA plate: one piece (96 wells / piece), sealed away from light, with desiccant inside.

[0164] Preparation of ELISA plate: Dilute the detection antigen to 3.125ug / mL with coating diluent (0.05mol / L, carbonate buffer solution with pH value of 9.6), 100μl per well, and put it in a 4℃ refrigerator overnight at 37℃ Coated for 2 hours, then blocked with 2% goat serum (200 μl / well) at 37°C for 2 hours. Wash with washing solution (PBS) 3 times, 3min / time.

[0165] ②Standard product: 7 bottles, the capacity of each bottle is 800μL.

[0166] Preparation of standard products: Add standard progesterone to it after hormone removal, the standard progesterone concentration is standard product 1 (20ng / mL), standard product 2 (10ng / mL), standard product 3 (5ng / mL), standard product 4 ( 2.5ng / mL), Standard 5 (1.25ng / mL), Standard 6 (0.625ng / mL), Standard 7 (0ng / m...

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Abstract

The invention relates to a detection kit of cow milk progesterone content which belongs to a detection kit. The kit contains progesterone monoclonal antibody which can perform the specific reaction with progesterone in dairy cattle. The kit is subject to strict inspections and can be used to measure the concentration of progesterone in a sample. The kit is the first independently developed cow milk progesterone detection kit in China, and the accuracy and minimum detectable quantity of the kit both meet the national standards.

Description

technical field [0001] The invention belongs to a detection kit, in particular to the detection of progesterone content in bovine milk. Background technique [0002] Progesterone is a hormone produced by the corpus luteum of the ovary during the estrous cycle (usually 21 days) of cows and cattle, and can enter blood, milk and saliva. Its concentration is lowest during the estrous period and reaches the highest level by the middle of the estrous cycle due to the maturation of the corpus luteum. [0003] Clinically, progesterone determination is mainly used to determine ovulation, monitor progesterone therapy, and evaluate early pregnancy status. With progesterone testing, it is possible to distinguish between pregnant cows, cows in estrus, or cows with irregular estrus cycles, so that appropriate measures can be taken quickly. The research on early pregnancy diagnosis technology of dairy cows has obvious economic significance for shortening calving interval, reducing insemi...

Claims

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Application Information

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IPC IPC(8): G01N33/74G01N33/577C07K16/26
Inventor 杨正涛冯晓声周晓菲张乃生
Owner JILIN UNIV
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