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Colloidal gold test strip for detecting four kinds of nitrofuran metabolites and preparation method thereof

A technology of colloidal gold test paper and nitrofuran, which is applied in the direction of biological testing, measuring devices, material inspection products, etc., can solve the problems of too many auxiliary equipment, the inability to achieve combined inspection of nitrofuran metabolites, and high requirements for the experimental environment. The effect of shortening the detection time, promoting the food safety cause, and improving the detection efficiency

Inactive Publication Date: 2016-12-21
北京艾旗斯德科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still deficiencies: (1) Laboratory instruments such as liquid chromatography, liquid chromatography-tandem mass spectrometer, and microplate reader are expensive and require a lot of initial investment and later maintenance costs; (2) sample processing and The detection requires a lot of auxiliary equipment, high requirements for the experimental environment, and a long detection cycle, which cannot monitor the source in time, and cannot meet the needs of a large number of nitrofuran metabolite surveys
However, a colloidal gold detection card can only detect one metabolite, and cannot achieve combined detection of more than four nitrofuran metabolites, and the detection efficiency is low

Method used

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  • Colloidal gold test strip for detecting four kinds of nitrofuran metabolites and preparation method thereof
  • Colloidal gold test strip for detecting four kinds of nitrofuran metabolites and preparation method thereof

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Experimental program
Comparison scheme
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Embodiment 1

[0023] 1. Measurement and preparation of colloidal gold: add 100mL ultrapure water and 1mL 1% chloroauric acid solution to the round bottom flask in the condensing reflux device, add 1mL 1% trisodium citrate solution after heating to boiling, and observe the color change. When the color is stable, continue to react for 10 minutes, cool and stand for a day, and use a particle size analyzer to obtain colloidal gold with a particle size of 40nm, and its maximum absorption peak is at 526nm.

[0024] 2. Preparation of colloidal gold-labeled antibody: the minimum amount of stable protein should be increased by about 10% for the optimal amount of labeled protein. Colloidal gold with 0.1mol / L of HCl and K 2 CO 3 Adjust the pH to 8.2, and label the antibodies separately. The method is as follows: Take the optimal amount of monoclonal antibody (the antibody labeling amounts of AOZ, AMOZ, AHD, and SEM are: 6 μg / mL, 8 μg / mL, 11 μg / mL, 9 μg / mL) 100mL of colloidal gold solution was added ...

Embodiment 2

[0031] 1. According to 1 in Example 1.

[0032] 2. Preparation of colloidal gold-labeled antibody: the minimum amount of stable protein should be increased by about 10% for the optimal amount of labeled protein. Colloidal gold with 0.1mol / L of HCl and K 2 CO 3 Adjust the pH to 8.2, and label the antibodies separately. The method is as follows: Take the optimal amount of monoclonal antibody (AOZ, AMOZ, AHD, and SEM antibody labeling amounts are: 5μg / mL, 7μg / mL, 9μg / mL, 8μg / mL) 100mL of colloidal gold solution was added to trehalose with a final mass concentration of 1%. After mixing for 10 minutes under a magnetic stirrer, BSA with a final mass concentration of 0.5% was added to obtain 100mL of each of the four gold-labeled antibodies, which were stored overnight at 4°C. Refrigerated ultracentrifugation for purification.

[0033] 3. According to the description in 3 of Example 1.

[0034] 4. According to the description in 4 of Example 1.

[0035] 5. According to the descr...

Embodiment 3

[0039] 1. According to 1 in Example 1.

[0040] 2. Preparation of colloidal gold-labeled antibody: the minimum amount of stable protein should be increased by about 10% for the optimal amount of labeled protein. Colloidal gold with 0.1mol / L of HCl and K 2 CO 3 Adjust the pH to 8.2, and label the antibodies separately. The method is as follows: Take the optimal amount of monoclonal antibody (the antibody labeling amounts of AOZ, AMOZ, AHD, and SEM are: 6 μg / mL, 8 μg / mL, 11 μg / mL, 9 μg / mL) 100mL of colloidal gold solution was added to trehalose with a final mass concentration of 1%. After mixing for 10 minutes under a magnetic stirrer, BSA with a final mass concentration of 0.5% was added to obtain 100mL of each of the four gold-labeled antibodies, which were stored overnight at 4°C. Refrigerated ultracentrifugation for purification.

[0041] 3. According to the description in 3 of Example 1.

[0042] 4. According to the description in 4 of Example 1.

[0043] 5. According to...

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Abstract

The invention relates to a colloidal gold test strip for detecting four kinds of nitrofuran metabolites and a preparation method thereof. The four kinds of nitrofuran comprise: furazolidone (AOZ), furaltadone (AMOZ), nitrofurantoin (AHD), and furacilin (SEM). The invention aims at providing the colloidal gold test strip for simultaneously detecting residues of four kinds of nitrofuran metabolites with fast and accurate characteristics and simple operation and the preparation method thereof. The aspects are improved, such as a method for marking four kinds of antibodies with colloidal gold, lineation on a nitrocellulose membrane (NC membrane), and treatment fluid for a combination pad; results show that the test strip has the advantages of high integration, high specificity, good repeatability, low cost, and the like. The operation is simple, the result is accurate, the method can be used for producing the test strip for combined detection of four kinds of nitrofuran metabolites, and the method has a certain guidance effect for producing test strips for combined detection.

Description

technical field [0001] The invention relates to the field of rapid detection of nitrofuran metabolites, in particular to a colloidal gold test strip for nitrofuran metabolites. Background technique [0002] Nitrofurans are widely used in livestock, poultry and aquaculture because of their low price and good effect to treat enteritis, scabies, red fin disease, canker, etc. caused by Escherichia coli or Salmonella. Because nitrofuran drugs and their metabolites have carcinogenic and teratogenic side effects on the human body, some countries have banned the use of nitrofuran drugs in livestock, poultry and aquatic animal foods, and strictly enforced the regulation of nitrofuran in aquatic products. Residue testing. Announcement No. 235 issued by the Ministry of Agriculture of the People's Republic of China on December 24, 2002 and Announcement No. 560 issued on October 28, 2005, nitrofuran drugs are prohibited in the feeding process. It shall not be detected in sexual food. ...

Claims

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Application Information

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IPC IPC(8): G01N33/94G01N33/558
CPCG01N33/94G01N33/558
Inventor 不公告发明人
Owner 北京艾旗斯德科技有限公司
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