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Colloidal gold test strip for detecting residue of sulfonamides, usage method and application thereof

A technology of colloidal gold test paper and sulfonamide drugs, which is applied in the field of veterinary drug residue analysis and immunology, can solve the problem of insufficient drugs, and achieves the effects of simple and rapid operation, easy wide-scale application and wide detection range.

Inactive Publication Date: 2014-06-25
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a colloidal gold test strip capable of detecting 16 kinds of sulfa drug residues at one time and its use method and application in view of the problem that the current sulfa drug residue detection test strip is not enough for one-time detection of drugs

Method used

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  • Colloidal gold test strip for detecting residue of sulfonamides, usage method and application thereof
  • Colloidal gold test strip for detecting residue of sulfonamides, usage method and application thereof
  • Colloidal gold test strip for detecting residue of sulfonamides, usage method and application thereof

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Embodiment 1

[0025] Embodiment 1 Preparation of colloidal gold test strips

[0026] 1.1 Preparation and purification of monoclonal antibodies

[0027] 1.1.1 Preparation of ascites: several Balb / c mice were taken 7 days before inoculation, and each mouse was pretreated by intraperitoneal injection of 0.5ml Freund's incomplete adjuvant. Use RPMI 1640 basal medium to suspend the cells expanded and cultured by the hybridoma cell 4E5 with the preservation number CCTCC NO:C201148, and adjust the number of cells to 1×10 6 cells / mL, each mouse was inoculated with 0.5ml intraperitoneally. Ascites was collected when the abdomen of the mice was obviously enlarged, the spirit became poor, and they were immobile when they were dying.

[0028] 1.1.2 Purification of monoclonal antibody: refer to the method in Zhu Liping's "Common Experimental Methods in Immunology" People's Military Medical Publishing House 2000 Edition: Take 5 ml of the obtained mouse ascites and mix it with an appropriate amount of s...

Embodiment 2

[0048] The using method and application of embodiment 2 colloidal gold test strip

[0049] 2.1 Optimization of colloidal gold test strip detection method

[0050] Dissolve the original coating, make 1mg / ml mother solution, dilute to 1mg / ml, 0.8mg / ml, 0.6mg / ml, 0.4mg / ml series concentration, use 10μg / L concentration of sulfamethoxine standard At the same time, use PBS detection as a negative control. When the drug detection line does not develop color at all and the negative control detection line develops obvious color, the coating concentration at this time is selected as the coating concentration of the colloidal gold test strip coating source.

[0051] Dilute goat or rabbit anti-mouse IgG to a concentration of 1mg / ml, 0.8mg / ml, 0.6mg / ml, 0.4mg / ml, 0.2mg / ml, and coat it on a nitrocellulose membrane as a quality control line. The color development of the quality control line and the detection line should be as consistent as possible, and the two detection lines of the negati...

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Abstract

The invention discloses a colloidal gold test strip, which comprises an absorption pad, a nitrocellulose membrane, a gold-labeled pad and a sample pad that are sequentially laminated and pasted on a plastic lining board. The cellulose nitrate membrane is provided with a detection line and a quality control line; the gold-labeled pad is coated with colloidal gold-labeled monoclonal antibodies capable of identifying sulfonamides; the detection line is coated with a sulfonamide coating antigen; and the quality control line is coated with a goat or rabbit anti-rat IgG. The invention also discloses a usage method of the colloidal gold test strip and application of the colloidal gold test strip to detection of residues of sulfonamides. The colloidal gold test strip provided by the invention can be used for detecting residues of sulfonamides in tissue samples, such as milk, pork, chicken, beef, lamb, fish, shrimp, pork liver, chicken liver, egg and honey, etc.; and the colloidal gold test strip provided by the invention can simultaneously detect 16 sulfonamides, and has characteristics of high sensitivity, strong specificity, simple operation, fastness, accuracy and low cost.

Description

technical field [0001] The invention belongs to the technical field of veterinary drug residue analysis and immunology, and in particular relates to a colloidal gold test strip for detecting sulfonamide drug residues and a use method and application thereof. Background technique [0002] Sulfonamide drugs are often widely used as feed additives or animal disease treatment drugs. Unreasonable drug use will lead to residues of sulfa drugs in animal foods, affecting food safety and endangering human health. More and more stringent regulations have been made on the residue limits of sulfa drugs in China. [0003] The detection methods for sulfa drug residues in food and feed include high performance liquid chromatography, gas chromatography, high performance liquid chromatography-mass spectrometry, gas chromatography-mass spectrometry, thin-layer chromatography, and capillary electrophoresis. Colloidal gold immunochromatography has the advantages of rapidity, high sensitivity, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577
Inventor 袁宗辉彭庆娟彭大鹏王玉莲陈冬梅陶燕飞刘振利
Owner HUAZHONG AGRI UNIV
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