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Immune colloidal gold test paper strip for detecting yapamicin relict and preparation thereof

A technology of colloidal gold test paper and kanamycin, which is applied in the field of immunochemical rapid detection of veterinary drug residues, can solve the problems of unfavorable promotion and use, complicated operation process, and cumbersome measurement methods, and achieve short detection time, low temperature requirements, The effect of easy operation

Inactive Publication Date: 2008-12-03
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chromatography or chromatography-mass spectrometry has high sensitivity and reliable results, but the determination method is cumbersome, time-consuming, and costly, and operators need professional training, so it is difficult to popularize
ELISA is a competitive immunoassay method using enzyme markers, which has been widely used as a rapid screening method for qualitative or semi-quantitative detection in recent years. Although the detection time has been shortened to a certain extent, it still requires specialized instruments (such as Microplate reader), the operation process is still complicated
More importantly, the above detection methods are all laboratory methods, which are difficult to apply in the field and are not conducive to popularization and use.

Method used

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  • Immune colloidal gold test paper strip for detecting yapamicin relict and preparation thereof
  • Immune colloidal gold test paper strip for detecting yapamicin relict and preparation thereof
  • Immune colloidal gold test paper strip for detecting yapamicin relict and preparation thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1: the structure of immune colloidal gold test strip of the present invention

[0031] Such as figure 1 and figure 2 As shown, an immunocolloidal gold test strip for detecting kanamycin residues, including sample pad 2, binding pad 3, nitrocellulose membrane 4, absorbent pad 5 and PVC backing 1, on PVC backing 1 There are sample pad 2, binding pad 3, nitrocellulose membrane 4 and water-absorbing pad 5 successively adhered on it. There is overlap between binding pad 3 and nitrocellulose membrane 4, and between There is overlap; the binding pad 3 is coated with anti-kanamycin monoclonal antibody-colloidal gold marker, and the nitrocellulose membrane 4 is coated with kanamycin-carrier protein (bovine serum albumin) coupling The detection line T composed of substances and the quality control line C composed of rabbit anti-mouse IgG. Finally cut into strips with a width of 3mm and vacuum package.

[0032] When in use, after a certain treatment of the sample, ...

Embodiment 2

[0034] Embodiment 2 (preparation embodiment)

[0035] Preparation method of immunocolloidal gold test strip for detecting kanamycin

[0036] l. Preparation of kanamycin-carrier protein conjugates

[0037] Synthesis of immunogen: Weigh 45 mg of kanamycin sulfate and 10 mg of hemocyanin (KLH) into 1.5 mL of PBS, and stir on a magnetic stirrer. Dissolve 300 mg of EDC·HCl in 1 mL of PBS, adjust the pH value to 7.4, and then add it dropwise to the above mixed solution, stir at room temperature for 1 hour, and then place it at 4°C for 3 days. Dialyze with PBS for 3 days, change the dialysate 2 to 3 times a day, pass through Sephadex G-25 for further purification, subpackage, and store at -20°C.

[0038] Synthesis of coating agent: Weigh 45 mg of kanamycin sulfate and 10 mg of BSA, dissolve in 1.5 mL of PBS, and stir on a magnetic stirrer. 300 mg of EDC·HCl was dissolved in 1 mL of PBS, and then added dropwise to the above mixed solution, and stirred and reacted at room temperatur...

Embodiment 3

[0057] Embodiment 3 (application embodiment)

[0058] Application Evaluation of Immunocolloidal Gold Test Strip for Detection of Kanamycin

[0059] 1. Specificity test

[0060] Prepare kanamycin, gentamicin, neomycin, streptomycin, chloramphenicol, cephalexin, sulfadimethoxine, and enrofloxacin into samples with a concentration of 10 ng / mL, and drop them on Test on the sample pad or on the sample well. Result of the test (seeing table 1) shows that the kanamycin sample detection line has no color, and a red band appears on the quality control line simultaneously, while gentamicin, neomycin, streptomycin, chloramphenicol, cephalexin , sulfadimethoxine, enrofloxacin sample test strip test line and quality control line all appear red bands, which indicates that gentamicin, neomycin, streptomycin, chloramphenicol, cephalexin, sulfa Dimethoxine and enrofloxacin have no cross-reaction with the test strip of the present invention, showing that the test strip of the present inventi...

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Abstract

The invention relates to a test strip used for detecting immune colloidal gold remaining in kanamycin. The test strip comprises a sample pad, a combination pad, a cellulose nitrate membrane, a water absorbing pad and a PVC back lining, wherein, the sample pad, the combination pad, the cellulose nitrate membrane and the water absorbing pad are continuously stuck on the PVC back lining in sequence,lapping is arranged between the combination pad and the cellulose nitrate membrane, and lapping is arranged between the cellulose nitrate membrane and the water absorbing pad; an anti-kanamycin monoclonal antibody-colloidal gold label is coated on the combination pad, and a detection line formed by a kanamycin-carrier protein (bovine serum albumin) coupling compound and a quality controlling lineformed by a rabbit anti mouse are coated on the cellulose nitrate membrane. The test strip of the invention has the advantages that the specificity is strong, the sensitivity is high, the detection time is short (10 minutes), the test strip can be operated on site, the detection cost is low, the operation is simple, and the test strip is not required to be operated by professionals.

Description

technical field [0001] The invention relates to an immunochemical rapid detection technology for veterinary drug residues, in particular to an immunocolloidal gold test strip for rapid detection of kanamycin residues and a preparation method thereof. Background technique [0002] Kanamycin is an aminoglycoside antibiotic that has significant antibacterial effects on a variety of Gram-positive and Gram-negative bacteria. It is widely used in animal husbandry as a feed additive and as an antibiotic for treating diseases. However, kanamycin has obvious toxic effects on human beings, and has serious damage to the cranial nerves, hearing and kidneys. In recent years, excessive residues in animal-derived foods due to the severe abuse of kanamycin have become one of the food safety issues that have attracted widespread attention at home and abroad. Many countries and international organizations have clearly stipulated the maximum residue limit (MRL) of the drug residue in food. F...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/558
Inventor 金涌邹明强陈立木王羚鸿郝刚
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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