Time resolved fluoro-immunochromatography method for quantitatively detecting beta-adrenoceptor agonists in animal

A time-resolved fluorescence and immunochromatography technology, applied in the field of immunology, can solve the problems of long time, complicated operation and high cost, and achieve the effect of improving solubility, good repeatability and high sensitivity

Inactive Publication Date: 2017-07-28
杭州泰熙生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Aiming at the disadvantages of complex operation, high cost and long time in the prior art, the present invention provides a simple, rapid and on-site time-resolved fluorescence immunochromatography method for quantitative detection of β-stimulants in animals

Method used

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  • Time resolved fluoro-immunochromatography method for quantitatively detecting beta-adrenoceptor agonists in animal

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Experimental program
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Effect test

Embodiment 1

[0027] A time-resolved fluorescent immunochromatography method for quantitative detection of beta agonists in animals, comprising the following steps,

[0028] Step one, Eu 3+ Labeled goat anti-mouse IgG: Take 5g / L goat anti-mouse IgG dissolved in 50mol / L, pH 7.0 PBS buffer, convert the buffer conditions through a PD-10 column, and the eluent is 0.05mol / L, pH 9.0. 6 carbonate buffer solution, dilute goat anti-mouse IgG to 1mg / mL with eluent, take 1.0mL diluted goat anti-mouse IgG and 0.5mg Eu 3+ - After DTTA was mixed, it was continuously stirred and reacted at 4°C for 24 hours. After the reaction, the resulting mixture was fractionated in a Sepharose CL-6B column using 0.05 mol / L, pH 8.0 Tris-HCl buffer solution containing 0.9% NaCl as the eluent, The fraction is Eu 3+ Labeled goat anti-mouse IgG;

[0029] Step two, Sm 3+ Labeled goat anti-rabbit IgG: Take 5g / L goat anti-rabbit IgG dissolved in 50mol / L, pH 7.0 PBS buffer, convert the buffer conditions through a PD-10 colu...

Embodiment 2

[0035] A time-resolved fluorescent immunochromatography method for quantitative detection of beta agonists in animals, comprising the following steps,

[0036] Step 1, take 5 g / L goat anti-mouse IgG dissolved in 50 mol / L, pH7.0 PBS buffer, and switch the buffer conditions through a PD-10 column, and the eluent is 0.05 mol / L, pH9.6 carbonic acid Saline buffer solution, dilute goat anti-mouse IgG to 1mg / mL with eluent, take 1.0mL diluted goat anti-mouse IgG and 0.5mg Eu 3+ - After DTTA was mixed, it was continuously stirred and reacted at 4°C for 24 hours. After the reaction, the resulting mixture was fractionated in a Sepharose CL-6B column using 0.05 mol / L, pH 8.0 Tris-HCl buffer solution containing 0.9% NaCl as the eluent, The fraction is Eu 3+ Labeled goat anti-mouse IgG;

[0037] Step two, Sm 3+ Labeled goat anti-rabbit IgG: Take 5g / L goat anti-rabbit IgG dissolved in 50mol / L, pH 7.0 PBS buffer, convert the buffer conditions through a PD-10 column, and the eluent is 0.05...

Embodiment 3

[0043] A time-resolved fluorescent immunochromatography method for quantitative detection of beta agonists in animals, comprising the following steps,

[0044] Step one, Eu 3+ Labeled goat anti-mouse IgG: Take 5g / L goat anti-mouse IgG dissolved in 50mol / L, pH 7.0 PBS buffer, convert the buffer conditions through a PD-10 column, and the eluent is 0.05mol / L, pH 9.0. 6 carbonate buffer solution, dilute goat anti-mouse IgG to 1mg / mL with eluent, take 1.0mL diluted goat anti-mouse IgG and 0.5mg Eu 3+ - After DTTA was mixed, it was continuously stirred and reacted at 4°C for 24 hours. After the reaction, the resulting mixture was fractionated in a Sepharose CL-6B column using 0.05 mol / L, pH 8.0 Tris-HCl buffer solution containing 0.9% NaCl as the eluent, The fraction is Eu 3+ Labeled goat anti-mouse IgG;

[0045] Step two, Sm 3+ Labeled goat anti-rabbit IgG: Take 5g / L goat anti-rabbit IgG dissolved in 50mol / L, pH 7.0 PBS buffer, convert the buffer conditions through a PD-10 colu...

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Abstract

The present invention relates to the field of immunology, and discloses a time resolved fluoro-immunochromatography method for quantitatively detecting beta-adrenoceptor agonists in an animal. The time resolved fluoro-immunochromatography method mainly comprises: (1) obtaining Eu<3+>-labeled goat anti-mouse IgG; (2) obtaining Sm<3+> labeled goat anti-rabbit IgG; (3) preparing a reference standard substance; (4) preparing a solid phase coated plate; and (5) taking the coated plate obtained in the step (4), adding 50 [mu]l of the standard substance obtained in the step (3) or a treated sample, adding 100 [mu]l of the label obtained in the step (1) and 100 [mu]l of the label obtained in the step (2), carrying out oscillation incubation for 30 min at a room temperature, washing five times with a 0.05 mol/L Tris-HCl washing liquid having the pH value of 7.4 and containing 0.9% of NaCl and 0.4-0.6% of methanol, adding 200 [mu]l of a reinforcing liquid, carrying out oscillation incubation for 10 min, and detecting with a semi-automatic fluorescence detector. The time resolved fluoro-immunochromatography method of the present invention has advantages of excellent stability, high sensitivity, and good reproducibility.

Description

technical field [0001] The invention relates to the field of immunology, in particular to a time-resolved fluorescent immunochromatography method for the quantitative detection of beta agonists in animals. Background technique [0002] Beta stimulant (beta adrenergic receptor agonist) drugs mainly include clenbuterol hydrochloride, ractopamine, salbutamol, bromobuterol, mabuterol, etc. Beta stimulants tend to accumulate in large quantities in the animal body and maintain Therefore, once the animal products are carried, it is difficult to remove them or make them inactivated, dissolved, and lost by general cooking methods and processes, thus seriously affecting human health. Countries such as Europe and the United States have long banned the use of β-stimulants in livestock and poultry production, and the Ministry of Agriculture of my country has also made the same regulations. However, driven by economic interests, illegal abuse of β-stimulants is still relatively common. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64G01N33/577
CPCG01N21/643G01N33/577G01N33/9413
Inventor 周斌王树明
Owner 杭州泰熙生物技术有限公司
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