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A kind of broad-spectrum detection card of glucocorticoid immunochromatography and its preparation method and application

A glucocorticoid and immunochromatography technology, applied in the field of detection, can solve the common problems of illegal addition, and achieve the effect of improving matrix interference.

Active Publication Date: 2022-03-29
深圳市计量质量检测研究院(国家高新技术计量站国家数字电子产品质量监督检验中心) +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the growth-promoting effect of glucocorticoids on animals and their efficacy in short-term whitening, illegal additions in meat-derived animal farming and cosmetics are common

Method used

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  • A kind of broad-spectrum detection card of glucocorticoid immunochromatography and its preparation method and application
  • A kind of broad-spectrum detection card of glucocorticoid immunochromatography and its preparation method and application
  • A kind of broad-spectrum detection card of glucocorticoid immunochromatography and its preparation method and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1 The synthesis of hapten for glucocorticoid immunization

[0037] Weigh 0.1 g of triamcinolone acetonide (or clobetasol, flumetasone), add 0.5 g of succinic anhydride respectively, adjust the pH to 4.5 after reacting at room temperature for 48 hours, filter the precipitate to obtain a precipitate, and name it as hapten 1, (or hapten 2 , hapten 3).

Embodiment 2

[0038] The synthesis of embodiment 2 glucocorticoid immune antigen

[0039] Weigh 0.1 g each of hapten 1, EDC, and NHS, add 2 mL of DMF, and stir for 24 hours. Add dropwise to 10mg / mL of KLH, continue stirring for 24 hours and then centrifuge. Glucocorticoid immune antigen 1 was obtained after the supernatant was dialyzed for 72 hours. Haptens 2 and 3 were coupled to KLH in the same way to obtain immune antigens 2 and 3.

Embodiment 3

[0040] Preparation and purification of embodiment 3 glucocorticoid antibody

[0041] New Zealand white rabbits were immunized with glucocorticoid immunization antigens 1, 2 and 3 respectively, 3 rabbits in each group. For the first immunization injection, 1 mL of 100 μg / mL immune antigen was fully emulsified with the same amount of Freund's complete adjuvant, and injected subcutaneously. After an interval of two weeks, take the sampled antigen, emulsify it with 1 mL of incomplete adjuvant, and inject it in the same way. After 4 times of immunization, blood was collected from the heart, and the serum was retained by centrifugation to obtain glucocorticoid antibodies 1, 2, and 3.

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Abstract

The invention provides a glucocorticoid immunochromatography broad-spectrum detection card as well as a preparation method and application thereof. It includes the following steps: step (1) synthesis of hapten for glucocorticoid immunization; step (2) synthesis of glucocorticoid immunization antigen; step (3) preparation and purification of glucocorticoid antibody; step (4) sugar Preparation of corticosteroid up-conversion luminescence-labeled antibody; step (5) preparation of coated antigen; step (6) preparation of glucocorticoid-coated antigen solid-phase nitrocellulose membrane; step (7) preparation of labeling pad; step (8) ) test strip assembly. The invention improves the detection range. The past glucocorticoid detection card can only detect several glucocorticoids such as dexamethasone and betamethasone. This invention combines three broad-spectrum antibodies due to the use of multiple up-converting chromogenic markers. on a test strip.

Description

technical field [0001] The invention belongs to the detection field, and in particular relates to a glucocorticoid immunochromatographic broad-spectrum detection card, a preparation method and application thereof. Background technique [0002] Glucocorticoids are a family of hormones containing more than 40 drugs. External use can reduce capillary permeability, reduce exudation and cell infiltration, and have powerful anti-inflammatory, anti-allergic, immunosuppressive and anti-proliferative effects. Therefore, it is widely used clinically in dermatology. In the breeding industry, it can promote the abnormal growth of animals and greatly improve the economic effect of animal breeding, so it is widely used in the breeding industry. However, the abuse of glucocorticoids will lead to varying degrees of residues in animal tissues and dairy products. After human consumption, it will lead to metabolic disorders and abnormal development of the body, and there are risks of carcinog...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/558G01N33/533
CPCG01N33/533G01N33/558
Inventor 张世伟姚添淇杨国武王士峰劳翠瑜冯荣虎林霖张恒李碧芳王坤吴佳辉
Owner 深圳市计量质量检测研究院(国家高新技术计量站国家数字电子产品质量监督检验中心)