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Gold labeled immunochromatographic strip assay semiquantitative detection test paper for detecting dienestrol

A semi-quantitative detection and dienestrol technology, used in measuring devices, biological testing, analytical materials, etc.

Inactive Publication Date: 2010-04-07
INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The enzyme-linked immunosorbent assay technology established based on this principle has the characteristics of convenience, rapidity, and sensitivity, but this technology requires testing personnel to have a high technical level, and also requires simple equipment such as a microplate reader, and the analysis time is generally 1-4 hours. , still has its limitation

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  • Gold labeled immunochromatographic strip assay semiquantitative detection test paper for detecting dienestrol
  • Gold labeled immunochromatographic strip assay semiquantitative detection test paper for detecting dienestrol

Examples

Experimental program
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Embodiment 1

[0031] The synthesis of embodiment 1 immunogen and the preparation of immune serum

[0032] 1.1 Preparation of Dienestrol Artificial Antigen:

[0033] Dienestrol conjugated antigens are divided into antigens for immunization and antigens for detection. The former is used for immunizing animals, and the latter is used for antibody detection and screening in antibody preparation and colloidal gold labeling or detection line coating in test paper preparation.

[0034] Antigen preparation for immunization:

[0035] Weigh 26.6 mg (0.1 mmol) of dienestrol and dissolve it in 1 ml of DMSO, stir thoroughly to dissolve it completely, add 11.7 mg of sodium chloroacetate, and stir at room temperature for 1 hour;

[0036] Weigh 50 mg of KLH, dissolve it in 0.1 mol / L PBS, slowly add DMSO dropwise and stir well to make it account for 50% of the total solution, add dienestrol activated by sodium chloroacetate to the KLH solution, add 10M hydrogen Adjust the pH value of the solution to 11 wi...

Embodiment 2

[0058] Embodiment 2 Preparation of dienestrol immunochromatography semi-quantitative detection test paper

[0059] 2.1 Sample liquid absorbent pad treatment:

[0060] Immerse the filter paper or glass fiber in 0.2mol / L PBS with pH7.4 for 2min, take it out, and dry it at 80℃ or by other methods to form a sample liquid absorption pad.

[0061] 2.2 Preparation and processing of colloidal gold marker pads:

[0062] The preparation and treatment of the colloidal gold labeling pad include the preparation of the colloidal gold sol, the colloidal gold-labeled bisestrol antibody, and the treatment of the colloidal gold labeling pad.

[0063] Colloidal gold sol preparation: HAuCl 4 Prepare 0.01% aqueous solution first, take 100ml and heat to boiling. Accurately add 1.2ml of 1% trisodium citrate (Na 3 C 6 h 5 o 7 2H 2 O) Aqueous solution. Continue heating and boiling for 15min. At this time, it can be observed that the light yellow chloroauric acid aqueous solution turns gray s...

Embodiment 3

[0068] Example 3 Dienestrol Gold-labeled Immunochromatography Semi-quantitative Detection Test Paper Sensitivity Experiment

[0069] Take standard solutions with concentrations of diethylestrol of 5ng / ml, 10ng / ml, and 15ng / ml respectively, prepare 10 prepared test strips for each concentration, insert the test strips into the above solution as required, and after 5 minutes Observation results.

[0070] The results showed that the detection lines on the test strips inserted into the 10ng / ml and 15ng / ml standard solutions were not visible, while the color of the detection lines on the test strips inserted into the 5ng / ml standard solution was not significantly different from the control line .

[0071] It can be seen from this that the dienestrol gold-labeled immunochromatographic semi-quantitative detection test paper of the present invention has high sensitivity, and the detection limit is 10 ng / ml.

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Abstract

The invention provides gold labeled immunochromatographic strip assay semiquantitative detection test paper for detecting dienestrol. The test paper is characterized in that a sample absorption pad, a colloid gold label pad, a chromatography pad and a water absorption pad are pasted on the back lining of the test paper; and a detection reaction pad is covered by dienestrol antigen serving as a detection line and second antibody serving as a quality control line at the same time. The semiquantitative detection test paper has the advantages that the test paper has strong specificity and can realize semiquantitative detection; the test paper can be used at a temperature of between 4 and 40 DEG C and can obtain a result after 5 minutes; and the test paper is suitable for quickly detecting dienestrol drugs in animal-derived foodstuff samples by sanitation departments, quality inspection departments, customs and livestock farms and foodstuff enterprises.

Description

Technical field: [0001] The invention relates to the technical field of immunoassay and veterinary drug residue detection. The invention relates to a rapid detection tool for estrogen dienestrol residues. Specifically, the invention relates to a gold-labeled immunochromatographic semi-quantitative detection test paper for dienestrol and its preparation method and use method. Background technique: [0002] Dienestrol (DE) is a synthetic non-steroidal estrogen, its chemical name is 3,4-bis(p-hydroxyphenyl)-2,4-hexadiene, which contains phenolic hydroxyl groups, It is a weakly acidic compound, which is mainly used to increase the milk production of dairy cows and the growth and fattening of livestock and poultry animals. Compared with natural estrogen, synthetic estrogen is usually more stable, stays in the body for a long time, and is easy to remain in fat and other tissues of humans and animals. Therefore, it can endanger human health through the food chain. The specific ma...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/74
Inventor 杨曙明于洪侠陈刚邱静张薇薇
Owner INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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