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Preparation method of colloidal gold test strip for quickly detecting chloramphenicol residues

A technology of colloidal gold test paper and chloramphenicol, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve problems such as the complex formula of the bonding pad treatment liquid, and achieve the effect of low price, good repeatability, and good stability

Inactive Publication Date: 2014-01-22
NANCHANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it is more complicated in the labeling method, the formulation of the resuspension solution during centrifugal purification, and the formulation of the binding pad treatment solution

Method used

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  • Preparation method of colloidal gold test strip for quickly detecting chloramphenicol residues
  • Preparation method of colloidal gold test strip for quickly detecting chloramphenicol residues

Examples

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Effect test

Embodiment 1

[0013] 1. Preparation of colloidal gold: add 100ml ultrapure water and 1ml 1% chloroauric acid solution to the round bottom flask in the reflux device, add 1ml 1% trisodium citrate solution after heating to boiling, observe the color change, Continue to react for 10 min when the color is stable. After cooling and standing for one day, the colloidal gold with a particle size of 40nm was detected by a particle size analyzer, and its maximum absorption peak was at 526nm.

[0014] 2. 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, take the optimal amount of chloramphenicol monoclonal antibody 16 μg / ml, trehalose with a final mass concentration of 1%, add 100ml of colloidal gold solution at the same time, mix for 10min under a magnetic stirrer, add the final mass concentration of 0.5% BSA, stored overnight at 4°C...

Embodiment 2

[0025] 1. According to 1 in embodiment 1.

[0026] 2. 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 9.2, take the optimal amount of chloramphenicol monoclonal antibody 16 μg / ml, trehalose with a final mass concentration of 0.5%, add 100ml of colloidal gold solution at the same time, mix under a magnetic stirrer for 10min, add the final mass concentration of 0.5% BSA, stored overnight at 4°C, then refrigerated and purified by ultracentrifugation.

[0027] 3. Purification of gold-labeled antibody:

[0028] (1) Centrifuge the gold-labeled antibody solution at room temperature and low speed (1500rpm) for 20min, discard the precipitate formed by the agglomerated gold particles, and take the red supernatant solution and centrifuge at 10000r / min for 30min at 4°C.

[0029] (2) The solution is divided into three layers...

Embodiment 3

[0037] 1. According to 1 in embodiment 1.

[0038] 2. 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 uses 0.1mol / L HCl and K 2 CO 3Adjust the pH to 9.2, take the optimal amount of chloramphenicol monoclonal antibody 16 μg / ml, trehalose with a final mass concentration of 2%, add 100ml of colloidal gold solution at the same time, mix for 10min under a magnetic stirrer, add the final mass concentration of 0.5% BSA, stored overnight at 4°C and purified by ultracentrifugation.

[0039] 3. Purification of gold-labeled antibody:

[0040] (1) Centrifuge the gold-labeled antibody solution at room temperature and low speed (1500rpm) for 20min, discard the precipitate formed by the agglomerated gold particles, take the red supernatant solution and centrifuge at 12000r / min for 30min at 4°C.

[0041] (2) The solution is divided into three layers: a transparent supernatant, ...

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Abstract

A preparation method of a colloidal gold test strip for quickly detecting chloramphenicol residues brings improvement to the method for labeling chloramphenicol antibodies with colloidal gold, heavy suspension and binding pad treatment solution aiming at the problem of complex conditions of the preparation processes of the prior art. The preparation process is simple and feasible. The results prove that the test strip has the advantages of high sensitivity and specificity, accuracy, simpleness, convenience and good repeatability. The method is stable, mature and reliable in the whole process conditions, can prepare the colloidal gold test strip with better quality and be used for producing the test strip for chloramphenicol and simultaneously has certain reference value to the production of other test strips.

Description

technical field [0001] The invention relates to a method for rapidly detecting chloramphenicol residues. Background technique [0002] Chloramphenicol (CAP) is an antibacterial agent produced by some specific Venezuelan bacteria in the soil, which has inhibitory effects on Gram-positive and negative bacteria, Rickettsia, Chlamydia, and Mycoplasma. However, it has been proved that its residues in animal foods will bring great toxic side effects to humans, such as aplastic anemia, and it is difficult to metabolize and eliminate. For this reason, the use of chloramphenicol has been banned in many countries, such as the United States, Canada, Australia and member states of the European Union. In 2002, the Ministry of Agriculture of China stipulated that chloramphenicol should not be detected in edible parts in the Regulations on Residues of Veterinary Drugs in Animal Food. However, due to the low price and good antibacterial effect of chloramphenicol, some criminals still use ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577
Inventor 刘成梅罗舜菁涂宗财陈文荣陈婷婷严杰琳高鹏陈臣
Owner NANCHANG UNIV
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