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A magnetic bead separation chemiluminescent immunodetection method for melamine

A melamine, magnetic separation technology, applied in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, measurement devices, etc., can solve the problems of human and animal health hazards, falsely increased protein, low acute toxicity, etc. The effect of stable luminescence value, low detection cost and wide detection range

Active Publication Date: 2018-05-04
CHENGDU CAPITALBIODX MEDICAL TECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The biggest feature of melamine is its high nitrogen content (66%), so it is often added to food and feed by unscrupulous traders to falsely increase the protein content, which has caused serious harm to human and animal health. Melamine has no metabolism in the organism Transformation, acute toxicity is very low, but long-term consumption will form urinary stones, causing kidney stones, kidney damage, renal failure, and even bladder cancer

Method used

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  • A magnetic bead separation chemiluminescent immunodetection method for melamine
  • A magnetic bead separation chemiluminescent immunodetection method for melamine
  • A magnetic bead separation chemiluminescent immunodetection method for melamine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Embodiment 1, melamine quantitative detection kit component and method thereof

[0069] 1. Preparation of melamine quantitative detection kit

[0070] The following 8 components are individually packaged and then packaged as a kit as a whole, which is the melamine quantitative detection kit: 1. Calibrator (containing a series of concentrations of MEL for establishing a standard curve); 2. Quality control product (containing A certain concentration of MEL); 3. Reagent A (containing a certain concentration of fluorescein isothiocyanate-labeled MEL antibody solution); 4. Reagent B (containing a certain concentration of alkaline phosphatase-labeled MEL antigen solution) 5. Sample Diluent (10mmol / L phosphate (PBS) buffer solution, the formula is the same as the calibrator buffer solution); 6. Magnetic separation reagent (magnetic bead suspension combined with anti-fluorescein isothiocyanate antibody); 7. Cleaning solution (for preparing magnetic bead cleaning solution); 8. ...

Embodiment 2

[0144] Embodiment 2, kit performance evaluation

[0145]According to the characteristics of in vitro diagnostic reagents, the linear range, sensitivity, accuracy and precision of the kit prepared in Example 1 were tested according to the usual practice. The specific operation steps are as follows:

[0146] 1. Reagent preparation:

[0147] Before the experiment, take out reagent A (the concentration of fluorescein isothiocyanate-labeled MEL monoclonal antibody is 0.1 μg / mL), reagent B (the concentration of alkaline phosphatase-labeled MEL antigen is 0.2 μg / mL), calibrator, Magnetic separation reagent, substrate solution, sample diluent, cleaning solution (obtained by mixing the cleaning solution concentrate and purified water at a ratio of 1:7), quality control products, and equilibrate to room temperature.

[0148] 2. Instrument preparation:

[0149] This kit uses ChemLite from Boao Bio Group Co., Ltd. TM 1200 automatic chemiluminescent immunoassay analyzer.

[0150] deta...

Embodiment 3

[0197] Embodiment 3, the application of kit

[0198] Sample addition recovery test: add 50ng / mL, 250ng / mL melamine standard substance respectively in the milk powder and feed that do not contain melamine, detect by the kit of one and two methods of embodiment 1, the milk powder recovery rate is 77.0%- 97.2%, feed recovery rate 78.0%-93.6%. The test data is shown in Table 5.

[0199] It can be seen that the kit and method of the present invention can detect melamine in the sample to be tested.

[0200] Table 5 Addition recovery

[0201]

[0202]

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Abstract

The invention discloses a quantitative detection kit for melamine. The invention provides a melamine quantitative detection kit, comprising the following 8 components: calibrator, quality control product, fluorescein isothiocyanate-labeled melamine antibody solution, alkaline phosphatase-labeled melamine antibody solution, sample dilution Liquid, magnetic separation reagent, cleaning solution, substrate solution; the detection method adopted in the present invention is magnetic bead separation chemiluminescence immunodetection, enzyme labeling technology, magnetic separation technology and chemiluminescence technology combined, its operation is convenient, simple, reaction The conditions are mild, the luminescence value is stable and less affected by external conditions. Compared with the existing detection methods, the invention has the advantages of simple sample processing process, low detection cost, rapid detection, accurate test results, good repeatability and the like.

Description

technical field [0001] The invention belongs to the technical field of immunological detection and analysis, relates to food safety-related immunological analysis and detection technology, and provides a magnetic bead separation chemiluminescence immunological detection method for melamine, which is suitable for the quantitative detection of melamine in samples such as milk, milk powder and feed. Background technique [0002] Melamine (Melamine, MEL), commonly known as melamine and protein essence, is a triazine nitrogen-containing heterocyclic organic compound, which is used as a chemical raw material and can be used in the production of plastics, coatings, adhesives, and food packaging materials. It is a white monoclinic crystal, almost odorless, slightly soluble in water, soluble in methanol, acetic acid, glycerin, etc., harmful to the body, and cannot be used in food processing or food additives. Those who artificially add melamine to food shall be investigated for legal...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/76G01N33/53G01N33/535
Inventor 韩学栋郭健夫高云邢婉丽
Owner CHENGDU CAPITALBIODX MEDICAL TECH CO LTD
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