Colloidal gold test paper strip for residue detection of tylosin and tilmicosin, method for using colloidal gold test paper strip and application of colloidal gold test paper strip

A technology of colloidal gold test paper and tylosin, which is applied in the field of veterinary drug residue analysis and immunology, can solve the problems of inconvenient on-site operation and complicated sample processing methods, and achieve small health hazards, fast detection speed, and simple sample processing methods Effect

Inactive Publication Date: 2012-10-03
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the test strip only involves the detection of a single drug, and the sample processing method provided by the invention is relatively complicated. When judging the test result, a strip reading instrument may be needed, which is not convenient for on-site operation.

Method used

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  • Colloidal gold test paper strip for residue detection of tylosin and tilmicosin, method for using colloidal gold test paper strip and application of colloidal gold test paper strip
  • Colloidal gold test paper strip for residue detection of tylosin and tilmicosin, method for using colloidal gold test paper strip and application of colloidal gold test paper strip
  • Colloidal gold test paper strip for residue detection of tylosin and tilmicosin, method for using colloidal gold test paper strip and application of colloidal gold test paper strip

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The preparation of embodiment 1 colloidal gold test strip

[0025] 1.1 Preparation and purification of monoclonal antibodies

[0026] 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 by the hybridoma cell line P3C4 with the preservation number CCTCC NO:C200719, 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.

[0027] 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 silic...

Embodiment 2

[0047] The using method and application of embodiment 2 colloidal gold test strip

[0048] 2.1 Optimization of colloidal gold test strip detection method

[0049] When the concentration of the coating source is greater than 0.05mg / ml, the test result of tylosin 10μg / L drug is negative; when the coating concentration is less than 0.05mg / ml, the test result of 10μg / L drug is positive, But the detection line of the blank sample is also very light in color. When the concentration of the coating source was 0.05 mg / ml, the result of detecting the concentration of tylosin 10 μg / L was positive, and the negative blank control test line developed a clear color.

[0050] When the quality control line goat or rabbit anti-mouse IgG coating concentration is 0.3mg / ml, the color of the negative blank quality control line is similar to that of the detection line. The final choice of goat or rabbit anti-mouse IgG coating concentration is 0.05mg / ml, and the concentration of the quality control...

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Abstract

The invention discloses a colloidal gold test paper strip. The colloidal gold test paper strip is formed by sequentially laminating an absorption pad, a nitrocellulose membrane, a gold label pad and a sample pad on a plastic lining plate, wherein a detection line and a quality control line is arranged on the nitrocellulose membrane; the gold label pad wraps a monoclonal antibody which has a colloidal gold label and can identify tylosin and tilmicosin; the detection line wraps desmycosin-O-carboxymethoxylamine-ovalbumin conjugates; and the quality control line wraps goat or rabbit anti-mouse immunoglobulin G (IgG). The invention also discloses a method for using the colloidal gold test paper strip and application of the colloidal gold test paper strip to residue detection of the tylosin and the tilmicosin. The invention can be applied to residue detection of the tylosin and the tilmicosin of tissues such as milk, chicken, pork, shrimp, beef, mutton and pork liver. The invention has the advantages of high sensitivity, high specificity, simplicity and convenience in operation, quickness and accuracy in detection, low detection cost and the like.

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 residues of tylosin and tilmicosin and its use method and application. Background technique [0002] Tylosin and tilmicosin are macrolide antibiotics, which are widely used in veterinary clinics to treat pneumonia, mastitis, and systemic sepsis caused by Gram-positive bacteria, especially for the treatment of mycoplasma infection in livestock and poultry . In addition, tylosin is also widely used as a feed additive. The residues of tylosin and tilmicosin in animal tissues will threaten food safety and cause harm to human health. Tylosin and tilmicosin can easily induce bacterial resistance, and the resistance can be transferred among bacteria. In view of this, the European Union banned the use of tylosin as a feed additive in 1999 (Council Regulation 2821 / 98, 1998), and my country's M...

Claims

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Application Information

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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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