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Colloidal gold test strip for semi-quantitative detection of progesterone and preparation method thereof

A colloidal gold test paper, semi-quantitative detection technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of expensive ELISA kits, difficulty in timely detection of non-pregnant cows, difficulty in widespread use in dairy farms, etc., and achieve improvement Reproductive performance, reduced empty time, control of obstetric diseases

Inactive Publication Date: 2011-12-28
TIANJIN AGRICULTURE COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Generally, artificial insemination must be carried out 2 months later, so it is difficult to find non-pregnant cows in time, and the cows cannot be treated in time, which increases the empty pregnancy time of cows
ELISA kits produced abroad are expensive, and it is difficult for domestic dairy farms to widely use them

Method used

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  • Colloidal gold test strip for semi-quantitative detection of progesterone and preparation method thereof
  • Colloidal gold test strip for semi-quantitative detection of progesterone and preparation method thereof
  • Colloidal gold test strip for semi-quantitative detection of progesterone and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0041] A colloidal gold test strip capable of detecting early-pregnancy cows, which consists of a glass cellulose membrane gold label pad 2 containing progesterone antibodies, a nitrocellulose membrane 7, a glass fiber membrane sample pad 1, a water-absorbing pad 6, and a PVC bottom plate 5 Developed by others, the test strip detection principle adopts the competition method.

[0042] The size of the test strip is 6cm×0.4cm, the length of the absorbent pad is 2cm, the length of the sample pad is 2cm, the length of the gold standard pad is 0.8cm, and the length of the nitrocellulose membrane is 2.5cm. Paste the nitrocellulose membrane on the base plate, 1.5cm away from the edge of the bottom plate’s water-absorbing end; Overlap in turn from top to bottom, the gold standard pad is located between the sample pad and the nitrocellulose membrane, and the overlapping length of the gold standard pad and the nitrocellulose membrane is 1 mm. The material of each material in this test ...

Embodiment 2

[0044] A preparation method of a colloidal gold test strip capable of detecting early pregnancy cows, the preparation steps are as follows:

[0045] The first step is to prepare colloidal gold by sodium citrate reduction method: first, heat 100mL 0.01% chloroauric acid solution to boiling in a microwave oven, then add 2.5mL 1% sodium citrate solution at one time, and mix quickly until the solution becomes Continue to boil for 3-5 minutes after turning into wine red. The diameter of the colloidal gold particles prepared in the present invention should be about 15nm-20nm.

[0046] The second step is to prepare and purify polyclonal antibody: Progesterone-11α-succinate (P 4 -11α-HS) were coupled with bovine serum albumin (BSA) and chicken ovalbumin (OVA) carriers to prepare progesterone artificial antigen P 4 -BSA and P 4 -OVA, respectively as the immunogen and the detection original (competition original). prepared P 4 -BSA and P 4 The concentrations of -OVA were 11.1 mg / m...

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Abstract

The invention relates to a colloidal gold test strip for semi-quantitative detection of progesterone and a preparation method thereof. The invention belongs to the technical field of dairy cow pregnancy diagnosis. Colloidal gold test strip for semi-quantitative detection of progesterone, the sample pad, the gold pad containing gold-labeled progesterone antibody complex, and the nitrocellulose membrane are pasted end to end on the bottom plate; there are detection lines and control lines on the nitrocellulose membrane . Preparation of test strips: preparation of colloidal gold: prepared from chloroauric acid and trisodium citrate; preparation of purified polyclonal antibody: preparation of artificial antigen of dairy cow progesterone by dicyclohexylcarbodiimide coupling method; artificial antigen of progesterone was then used Immune mice to prepare progesterone polyclonal antibody; prepare gold-labeled antibody: colloidal gold and antibody adsorption, centrifugal purification; colloidal gold test strip preparation: soak gold-labeled pad with Tween-containing PB solution, gold-labeled antibody perfusion glass fiber Membrane; nitrocellulose membrane Spray bovine progesterone artificial antigen and goat anti-mouse secondary antibody into 2 lines with a film dispenser. The invention is simple in operation, convenient and practical, efficient and fast, and easy to popularize.

Description

technical field [0001] The invention belongs to the technical field of dairy cow pregnancy diagnosis, in particular to a colloidal gold test strip for semi-quantitative detection of progesterone and a preparation method thereof. Background technique [0002] Progesterone (P 4 ) is mainly synthesized in the ovary, which is an accurate indicator reflecting the activity of the ovary, and has the function of controlling the reproductive activities of female animals. Detecting the progesterone level and content changes in body fluids can not only understand the reproductive physiology of female animals, but also monitor reproductive system diseases. an effective method. P in blood or milk 4 The determination of its content can also be used for early pregnancy diagnosis, infertility diagnosis and estrus identification; it can also be used for research on ovarian function, reproductive disorders and obstetrical diseases; it can also be used to check the effect of artificial inse...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/544
Inventor 杨百亮李志源艾霞张建斌李天俊
Owner TIANJIN AGRICULTURE COLLEGE
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