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Colloidal gold test strip for testing melamine content

A technology of colloidal gold test paper and melamine, which is applied in the directions of measuring devices, biological tests, material inspection products, etc., can solve the problems of low specificity and high cost, and achieve the effects of improving specificity, reducing cost and improving visibility.

Inactive Publication Date: 2009-09-23
李红玉 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current ELISA melamine detection kit based on the principle of immunology still cannot get rid of the shackles of the instrument, and the cost is also high. It takes 1-2 hours, and it uses polyclonal antibodies, which is not specific. high

Method used

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  • Colloidal gold test strip for testing melamine content
  • Colloidal gold test strip for testing melamine content

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] The preparation of embodiment 1 anti-melamine monoclonal antibody

[0018] (1) Carboxylate melamine into carboxyl melamine with succinic acid, then couple it with hemocyanin KLH and chicken ovalbumin OVA by carbodiimide method, purify melamine-coupled protein by dialysis, scan with ultraviolet method to identify its coupling effect. Finally, complete melamine antigen (immunization antigen MEL-KLH, coating antigen MEL-OVA) was made.

[0019] (2) BALB / C mice were immunized with melamine immune antigen MEL-KLH. After three immunizations, the serum antibody titer of the mice was measured. After booster immunization, the mouse with the highest titer was selected as the splenocyte provider.

[0020] (3) The SP2 / 0 cells were domesticated with 8-azaguanine, and after they were domesticated, the SP2 / 0 cells were fused with mouse splenocytes that produced high antibody levels in vivo according to the classical animal cell fusion method.

[0021] (4) Then use the melamine-coated...

Embodiment 2

[0024] The preparation of embodiment 2 mouse anti-chicken ovalbumin (OVA) antibody

[0025] (1) BALB / C mice were immunized with chicken ovalbumin according to the routine immunization procedure, and the mouse antibody titer was determined by ELISA method after three immunizations to determine the mouse antibody level.

[0026] (2) The mouse with the highest titer was subjected to enucleation to collect blood, and its serum was separated to obtain its serum, which was purified by octanoic acid-ammonium sulfate method to obtain the mouse anti-chicken ovalbumin OVA antibody.

Embodiment 3

[0027] The preparation of embodiment 3 colloidal gold nanoparticles

[0028] (1) Add a certain amount (100mL) of 0.01% chloroauric acid aqueous solution into a clean beaker, then heat to boiling.

[0029] (2) Add 1 mL of trisodium citrate aqueous solution with a certain concentration (1%) under stirring state, and continue to boil for 10 minutes, the color of the reaction system gradually changes from golden yellow to purple.

[0030] (3) After standing to cool, restore to the original volume with deionized water, and finally make qualified (about 40 nm in diameter) colloidal gold nanoparticles.

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Abstract

The invention relates to a colloidal gold test strip for testing melamine content. The test strip mainly comprises a supporting plate, a sampling zone, a colloidal mark redissolve zone, a color development zone, a water absorbing pad and the like. By adopting the theory of competition method, melamine in a sample is competed with gold mark melamine envelope antigen MEL-OVA, therefore the melamine in the sample is combined with anti-melamine monoclonal antibody on the test strip so as to be developed, and the corresponding relation of the melamine content in the sample and the color development quantity of the test strips is built, therefore the melamine content can be judged according to the color development quantity of the strips in the color development zone. The colloidal gold test strip has the advantages of high specificity, high sensitivity, fast testing speed, simple operation, convenient portability and the like, and can quick and sensitively judge whether the samples, such as milk, feed, animal product, blood and urine, contains melamine, or whether the melamine content exceeds the standard, thus presenting evaluation and judgment to food security and human body health. The colloidal gold test strip can be suitable to the on-site quick screening of food, feed and the like, and can also be applied to the self-check in the daily life at home and autologous melamine content testing of human.

Description

1. Technical field [0001] This invention is a colloidal gold test strip for detecting the content of melamine, specifically a semi-quantitative colloidal gold rapid detection test strip for melamine, which is mainly used for personal health, food, feed safety and animal products. The detection of melamine content. It belongs to the field of monoclonal antibody technology, gold-labeled antibody technology and competition immunochromatography technology. 2. Background technology [0002] Melamine is a triazine-type nitrogen-containing heterocyclic organic compound and an important nitrogen-heterocyclic organic chemical raw material. Widely used, it is a basic organic chemical intermediate product. [0003] Melamine is banned from food and animal feed because it forms insoluble melamine cyanurate in the human body, causing serious kidney stones. [0004] In 2007, an incident of contaminated pet food broke out in the United States, causing some pet cats and dogs to suffer sev...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/558G01N33/566G01N33/52
Inventor 李红玉徐志良
Owner 李红玉
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