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Melamine colloidal gold test card as well as production and application method thereof

A technology of melamine and a production method, which is applied in the field of immunology and achieves the effects of simple operation, low detection cost and stable results

Active Publication Date: 2011-04-06
深圳市三方圆生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

And there is no report of this technology monitoring melamine both at home and abroad

Method used

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  • Melamine colloidal gold test card as well as production and application method thereof
  • Melamine colloidal gold test card as well as production and application method thereof
  • Melamine colloidal gold test card as well as production and application method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1 3

[0043] The synthesis of embodiment 1 melamine hapten

[0044] Weigh 0.60 g of 4-aminobutyric acid and 0.4 g of NaOH, dissolve them in 20 mL of water, and slowly add an equivalent amount of 6-chloro-2,4-diamino-1,3,5-triazine at room temperature. Gradually raise the temperature to reflux state, and react for 4h. Adjust the pH to 6, and there is precipitation. The precipitate was filtered and washed twice with distilled water to obtain 0.5 g of melamine hapten.

[0045] The prepared hapten structure is:

[0046]

Embodiment 2 3

[0047] Synthesis of embodiment 2 melamine immune antigen

[0048] Take melamine hapten 0.1mmol and dissolve in 2mL DMF, stir and add 27.5mg DCC and 14.4mg NHS. Magnetic stirring was carried out overnight at 4°C. After centrifugation, the supernatant was liquid A, and 140 mg of BSA was weighed and dissolved in 10 mL of PBS (pH 8.0) with a concentration of 0.1 mol / L. Add 1 mL of DMF, stir and dissolve to prepare liquid B. Under magnetic stirring, liquid A is gradually dropped into liquid B, and react at 4°C for 12 hours. After centrifugation, the supernatant was taken, dialyzed with normal saline at 4°C for 3 days, and the dialysate was changed 3 times a day. The obtained whole antigen was dispensed into 0.5 mL centrifuge tubes at a concentration of 1 mg / mL. Store in -20°C freezer.

[0049] The prepared immune antigen structure is:

[0050]

Embodiment 3

[0051] Embodiment 3 experimental animal immunization

[0052] The hapten melamine immune antigen was used to immunize female balb / c mice for 6 weeks respectively, 3 mice in each group. For the first immunization injection, 100 μL of 100 μg / mL immune antigen was fully emulsified with the same amount of complete Freund’s adjuvant, and injected directly into the intraperitoneal cavity. After an interval of two weeks, take the sampled antigen, emulsify it with 100 μL of incomplete adjuvant, and inject it in the same way.

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Abstract

The invention relates to a melamine colloidal gold test card as well as a production and application method thereof, belonging to the technical field of immunology. The rapid diagnosis card comprises a shell and a test paper strip and is characterized in that the shell is provided with a sample application hole and an observation window, the test paper strip is arranged in the shell, a cellulose nitrate film is superposed in the middle part of a polyvinyl chloride (PVC) liner plate, a water-absorbing pad and a sample pad are superposed at both ends of the PVC liner plate, the cellulose nitrate film overlaps with the water-absorbing pad and the sample pad, a length of film containing a melamine monoclonal antibody immune colloidal gold compound is attached to a sample area, and the cellulose nitrate film is provided with a test tape containing melamine coating antigens and a quality control tape containing goat-anti-mouse IgG. The test card of the invention is a rapid, accurate and sensitive diagnoses card, does not need any special instrument, has the advantage of simple operation and can meet the test requirements of food safety, animal husbandry and testing organization.

Description

technical field [0001] The invention relates to a melamine colloidal gold detection card, which belongs to the technical field of immunology. Background technique [0002] Melamine (English name: Melamine) is a triazine-type nitrogen-containing heterocyclic organic compound, an important nitrogen-heterocyclic organic chemical raw material. Due to the flaws in the protein content testing methods of the food and feed industry, melamine is often used as a food additive by unscrupulous businessmen to improve the protein content indicators in food testing, so melamine is also known as "protein essence". [0003] Proteins are mainly composed of amino acids. The average nitrogen content of protein is about 16%, while the nitrogen content of melamine is about 66%. The commonly used protein testing method "Kjeldahl method" is to estimate the protein content by multiplying the measured nitrogen content by 6.25. Therefore, the addition of melamine will make the protein test content o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577C07K14/765C07K14/77C07D251/70
Inventor 钟松清谭攀李细清张世伟
Owner 深圳市三方圆生物科技股份有限公司
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