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Protein chip for quickly detecting veterinary medicinal residue in food and agricultural products

A technology for agricultural and sideline products and veterinary drug residues, applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of weak specificity, complex operation, and low sensitivity of protein chips.

Inactive Publication Date: 2009-11-18
台州金芯生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the disadvantages of weak specificity, low sensitivity, narrow application range and complicated operation of the protein chip used in the prior art, the present invention provides a chip with simple structure, convenient operation, low price, continuous, integrated and miniaturized detection and analysis process. , High-throughput veterinary drug residue detection protein chip

Method used

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  • Protein chip for quickly detecting veterinary medicinal residue in food and agricultural products
  • Protein chip for quickly detecting veterinary medicinal residue in food and agricultural products
  • Protein chip for quickly detecting veterinary medicinal residue in food and agricultural products

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Preparation of protein chip

[0039] Spot 2μl of the antigen solution diluted in 0.005M PBS pH7.2 on the nitrocellulose membrane, dry it in a 37°C incubator for 20-30 minutes, add the blocking solution dropwise in a 37°C incubator to block for 10 minutes, and wash with washing solution for 1-30 minutes. 2 times, filter paper blotted dry.

[0040] Wherein the solid phase carrier is: nitrocellulose membrane; the veterinary drugs used on the solid phase carrier are: chloramphenicol, streptomycin, tetracycline.

[0041] Diluent: 0.005M PBS pH7.2.

[0042] Blocking solution and its concentration: 5% skimmed milk.

[0043] Washing solution: 0.01M PBS pH7.4 containing 0.5% Tween20,

[0044]0.2M Na2HPO4 87ml

[0045] 0.2M NaH2PO4 13ml

[0046] NaCl 15.2g

[0047] Tween 206ml

[0048] Add double steamed H 2 0 to 2000ml.

[0049] Dissolve the conjugated antigens of veterinary drugs to be prepared on the nitrocellulose membrane in the coating solution at a concentration o...

Embodiment 2

[0052] Preparation of protein chip

[0053] Spot 2 μl of the antigen solution diluted in 0.01M PBS pH7.4 on the nitrocellulose membrane, dry it in a 37°C incubator for 20-30 minutes, add blocking solution dropwise in a 37°C incubator to block for 10 minutes, and wash with washing solution for 1-30 minutes. 2 times, filter paper blotted dry.

[0054] Wherein the solid phase carrier is: nitrocellulose membrane; the veterinary drugs used on the solid phase carrier are: chloramphenicol, streptomycin, tetracycline.

[0055] Diluent: 0.01M PBS pH7.4.

[0056] Blocking solution and its concentration: 5% skimmed milk.

[0057] Washing liquid is the same as embodiment 1

[0058] Dissolve the conjugated antigens of veterinary drugs to be prepared on the nitrocellulose membrane in the coating solution at a concentration of 0.8 mg / ml, and then use the automatic chip spotting system to apply these veterinary drugs according to the amount used at 1.2 μg / point, and the spotting density M...

Embodiment 3

[0060] Preparation of protein chip

[0061] Spot 2 μl of antigen solution diluted in 0.02M PBS pH7.6 on the nitrocellulose membrane, dry in a 37°C incubator for 20-30 minutes, add blocking solution dropwise in a 37°C incubator to block for 10 minutes, and wash with washing solution for 1-30 minutes 2 times, filter paper blotted dry.

[0062] Wherein the solid phase carrier is: nitrocellulose membrane; the veterinary drugs used on the solid phase carrier are: chloramphenicol, streptomycin, tetracycline.

[0063] Diluent: 0.02M PBS pH7.6.

[0064] Blocking solution and its concentration: 5% skimmed milk.

[0065] Washing liquid is the same as embodiment 1

[0066] Dissolve the conjugated antigen of veterinary drugs to be prepared on the nitrocellulose membrane in the coating solution at a concentration of 0.8 mg / ml, and then use the chip automatic spotting system to apply these veterinary drugs according to the amount used at 0.8 μg / point, and the spotting density 25 dots / cm...

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Abstract

The invention provides a method for detecting residues of veterinary drugs (products) in agricultural by-products and food, and belongs to the technical field of toxic substance detection. The detection method for rapid detection of veterinary drug (product) residues in food and agricultural by-products comprises the following steps: spotting multiple veterinary drug (product)-coupled antigens on a solid phase carrier according to the microarray and coating it; wherein the solid phase The veterinary drug (product) antigen is immobilized on the matrix; the biotinylated antibody diluted with the diluent is mixed with the sample to be tested, and a competitive immune reaction is performed with a variety of veterinary drug (product) coated antigens immobilized on the solid phase carrier, and the washing solution Washing; add avidin-labeled colloidal gold for amplification and staining, and obtain grayscale results through a scanner. Compared with the detection of the prior art, the invention adopts the immunocompetition method to detect, and mainly has the characteristics of high sensitivity, strong specificity and easy operation. The immunocompetitive method requires fewer reagents, simple sample addition procedure, and short reaction time; at the same time, measurement and data processing are easy to automate.

Description

technical field [0001] The invention belongs to the technical field of biochips, in particular to a protein chip used for detecting veterinary drug residues in food and agricultural by-products. Background technique [0002] my country is a big country in the production and consumption of agricultural products, and the detection of veterinary drug residues in agricultural products is directly related to the production, export and food consumption safety of agricultural products in my country. In particular, the export of agricultural products in my country has been subject to foreign technical barriers to trade in recent years due to veterinary drug residues. Only the export of agricultural products in Zhejiang Province has been banned for hundreds of millions of dollars a year. In recent years, food safety issues have become one of the key issues of national concern. Food pollution incidents and export obstructions caused by toxic residues exceeding the standard have occurr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N33/531
Inventor 王华全林荣业
Owner 台州金芯生物工程有限公司