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Immune colloidal gold strip for detecting residue of roxarsone and preparation method thereof

A technology of roxars arsenic and test strips, which is applied in the field of immunochemical rapid detection of veterinary drug residues, can solve the problems of complex operation process, high detection cost, expensive equipment, etc., and achieve low temperature requirements, short detection time, and high detection efficiency. low cost effect

Inactive Publication Date: 2010-07-14
DALIAN PRACTICAL BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chromatographic technology is very effective and accurate for the detection of roxarsarsenic, but it also has the following disadvantages: (1) the pretreatment procedure of the sample to be tested is cumbersome and time-consuming; Relevant experience, operators must understand the various interference factors that affect chromatographic analysis, understand the advantages and disadvantages of the method used, in order to obtain reliable analysis results; (3) need expensive equipment to assist, it is difficult to popularize in small and medium-sized enterprises; (4) ) The requirements for instrument maintenance are high, and the quality of maintenance directly affects the accuracy of the analysis results; (5) The detection cost is high
The disadvantages are: (1) special equipment such as a microplate reader is required for use, which is not conducive to popularization and use in grassroots units; (2) detection operators need to undergo professional training; (3) the operation process is relatively complicated, The detection takes a long time, the whole process takes 2-4h

Method used

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  • Immune colloidal gold strip for detecting residue of roxarsone and preparation method thereof
  • Immune colloidal gold strip for detecting residue of roxarsone and preparation method thereof
  • Immune colloidal gold strip for detecting residue of roxarsone and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1 (preparation embodiment)

[0033] Preparation method of immunoassay strip for detecting roxarsarsenic

[0034] 1. Preparation of roxarsarsenic-carrier protein conjugates

[0035] Synthesis of roxarsarsenic-carrier protein conjugates:

[0036] (1) Synthesize ROX-HSA whole antigen, the specific steps are as follows: first add 20mg ROX to 300μl dimethylformamide solution, then slowly add to 1ml 0.5mol / LH 2 SO 4 , then 1ml 1mol / L NaNO was mixed under magnetic stirring 2 Slowly added to the SMZ solution and reacted for 10 minutes to form a diazonium salt.

[0037] (2) Under magnetic stirring, 100 mg of HSA and 100 mg of OVA were respectively added to 4 ml of phosphate buffer.

[0038] (3) The diazotization salt solution formed above was slowly added to the HSA solution and the OVA solution under magnetic stirring, and the pH was maintained at 8-10 with 1M NaOH, and reacted at 4° C. for 1 h.

[0039] (4) The reaction solution was dialyzed in normal saline f...

Embodiment 2

[0059] Embodiment 2 (application embodiment)

[0060] Method for using immunocolloidal gold test strip for detecting roxarsarsenic

[0061] 1. Sample pretreatment

[0062] (1) Pretreatment of pig urine

[0063] Centrifuge 2ml of the test urine at 2000rpm for 10min, and take the supernatant for testing.

[0064] (2) Pretreatment of milk samples

[0065] The milk sample was centrifuged at 3500rpm for 10min, 3ml was taken and added with 6ml ethyl acetate and shaken up and down for 10min, centrifuged at room temperature 3000rpm for 10min, 2ml of the upper liquid was evaporated to dryness or dried in a nitrogen stream, and 1ml of 0.01M pH7.4 phosphate buffer ( PBS) to dissolve the residue.

[0066] (3) Pretreatment of tissue samples

[0067] Homogenize the tissue sample to be tested, take 3g of the tissue sample to be tested, add 6ml of ethyl acetate, shake up and down for 10min, centrifuge at 3000rpm at room temperature for 10min, take 2ml of the upper liquid and evaporate it...

Embodiment 3

[0074] Embodiment 3 (application embodiment)

[0075] Application of Immunocolloidal Gold Test Strips for Detection of Roxars Arsenic

[0076] 1. Specificity test

[0077] The roxarsarsenic, nifelic acid and arsanic acid were prepared into samples with a concentration of 600ppb. Test according to the method described in Example 2. The test results (see Table 1) show that there is no color on the detection line of the Roxarsarsenic sample, and a red band appears on the quality control line simultaneously, while the color of the detection line of the test paper of the nifedipine and arsanic acid samples is the same as that of the standard product test paper. The colors of the test lines are consistent, and red bands appear on the quality control line simultaneously, which shows that nifeterine, arsanic acid and the test strips of the present invention have no cross-reaction, showing that the test strips have good specificity.

[0078] Table 1 test strip specificity test resul...

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Abstract

The invention discloses an immune colloidal gold strip for detecting residue of roxarsone and a preparation method thereof, and belongs to the technical field of immunochemistry quick test for residue of a veterinary drug. The strip comprises a sample pad, a combined pad, a nitrocellulose membrane, a water absorption pad and a PVC back lining; and the immune colloidal gold strip is characterized in that: the PVC back lining is adhered with the sample pad, the combined pad, the nitrocellulose membrane and the water absorption pad in sequence; the combined pad is coated with an anti-roxarsone monoclonal antibody-colloidal gold marker; and the monoclonal antibody is obtained by excretion of a hybridoma cell line. The nitrocellulose membrane is coated with a detection line consisting of a roxarsone-carrier protein conjugate and a quality control line consisting of rabbit anti-mouse IgG. The strip of the invention has remarkable advantages of high sensitivity, strong specificity, simple and convenient operation, rapid detection, accuracy and the like.

Description

technical field [0001] The invention belongs to the technical field of immunochemical rapid detection of veterinary drug residues, specifically, an immunochromatographic reaction using colloidal gold markers for color development, an immunocolloidal gold test strip for rapid detection of roxarsine residues and its preparation method. Background technique [0002] In recent years, roxarsarsenic has been widely used as a feed additive in animal husbandry because of its pharmacological properties of stimulating animal growth, anti-parasite and broad antibacterial spectrum. [0003] Although the toxicity of roxarsarsenic is low, it can still cause poisoning when used in large doses, such as vitamin B1 deficiency, muscle tremors, ataxia and even death. In addition, it will cause great pollution to the environment. If a large amount of organic arsenic preparations are used in livestock and poultry diets, the arsenic-containing livestock and poultry excrement will enter the enviro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
Inventor 刘汉石
Owner DALIAN PRACTICAL BIOTECH
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