Immune colloidal gold test strip for detection of ractopamine residue in swine urine

A technology of ractopamine and colloidal gold, which is applied in the field of immunochemical rapid detection of veterinary drug residues, can solve the problems of complex operation, long detection time, high detection cost, etc., and achieve the effect of low temperature requirements, low detection cost and short detection time

Inactive Publication Date: 2013-06-19
DALIAN PRACTICAL BIOTECH
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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 ractopamine, but it also has the following disadvantages: (1) The pretreatment procedure of the sample to be tested is cumbersome and time-consuming; (2) It needs professionally trained technicians to operate, and the operators must have rich experience , it is necessary to understand the various interference factors affecting chromatographic analysis and the advantages and disadvantages of the method used in order to obtain reliable analysis results; (3) it requires expensive equipment and equipment, which is difficult to popularize in small and medium-sized enterprises; (4) the maintenance of the instrument The requirements are high, and the quality of maintenance directly affects the accuracy of the analysis results; (5) The detection cost is high
However, there are still the following disadvantages: (1) special instruments and equipment such as microplate readers are required to cooperate with the use, which is not conducive to the promotion and use of grassroots units; (2) detection operators need to undergo professional training; (3) the operation is more complicated and the detection time Longer, the whole process takes 2-4 hours

Method used

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  • Immune colloidal gold test strip for detection of ractopamine residue in swine urine
  • Immune colloidal gold test strip for detection of ractopamine residue in swine urine
  • Immune colloidal gold test strip for detection of ractopamine residue in swine urine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The preparation method of embodiment 1 ractopamine detection test paper

[0030] 1. Coupling of Ractopamine RAC to Carrier Protein BSA

[0031] Rac is coupled with carrier protein by diazotization method to prepare immune antigen. Dissolve 5mg of ractopamine in pre-cooled (2-8°C) dilute hydrochloric acid (pH3.0), add 10mg of sodium nitrite (weight ratio: 5:1), stir at room temperature in the dark for 6 hours, the color of the solution turns yellow . The carrier bovine serum albumin was dissolved in PBS buffer, added to the above reaction at a molar ratio of 25:1, and stirred overnight in the greenhouse. The reaction solution was dialyzed against PBS buffer.

[0032] 2. Preparation of anti-ractopamine Rac monoclonal antibody (W1)

[0033] Immunize BALB / c mice with 50ug-100ug / carrier protein-coupled Rac for three times, with an interval of 15-30 days between each time; after the third booster immunization, 3-4 years old, the immunized mice were bled from the eyes and ...

Embodiment 2

[0041] Ractopamine rapid detection test strip detection operation method

[0042] 1. Sample pretreatment

[0043] Pretreatment of Pig Urine

[0044] Centrifuge 2ml of the test urine at 2000rpm for 10min, and take the supernatant for testing.

[0045] 2. Detection

[0046] Take the sample to be tested with a dropper, drop 3 drops in the sample hole of the test card, and observe the result after 5 minutes.

[0047] 3. Result judgment

[0048] like image 3 As shown, if the test strip test line and the quality control line of the sample to be tested appear red bands at the same time, it is judged as a negative sample, that is, the concentration of ractopamine in the sample to be tested is lower than 3ppb (such as image 3 -A shown); if no color appears on the test strip detection line of the sample to be tested, and a red band appears on the quality control line at the same time, it is judged as a positive sample, that is, the concentration of ractopamine in the sample to be...

Embodiment 3

[0050] Application of immunocolloidal gold test strips for detecting ractopamine

[0051] 1. Specificity test

[0052] Clenbuterol, albuterol, tobuterol, and terbutaline were prepared into samples with a concentration of 5ppb. Test according to the method described in Example 2. The test results (see Table 1) show that the ractopamine sample detection line has no color, and a red band appears on the quality control line simultaneously, while clenbuterol, albuterol, tobuterol, and terbutaline sample test strips detect The color of the line is consistent with the color of the detection line of the standard product test strip, and a red band appears on the quality control line simultaneously, which shows that clenbuterol, albuterol, tobuterol, terbutaline and the test strip of the present invention do not cross The reaction shows that the test strip has good specificity.

[0053] Table 1 test strip specificity test result of the present invention

[0054] Test sample...

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Abstract

The present invention discloses an immune colloidal gold test strip for the rapid detection of ractopamine residues in animal urine and a preparation method thereof, belonging to the technical field of immunochemical fast detection of residues of veterinary drugs and forbidden drugs. The test strip (shown in Figure 1) of the present invention includes a sample pad (1), a colloidal gold combination pad (2), a nitrocellulose membrane (3), an water absorption pad (4) and a PVC backing (7), and is characterized in that the sample pad (1), the combination pad (2), the nitrocellulose membrane (3) and the water absorption pad (4) are adhered on the PVC backing (7) in sequence; and monoclonal antibody of ractopamine-colloidal gold markers are coated on the combination pad (2), wherein the monoclonal antibody is secreted by a hybridoma cell line. The nitrocellulose membrane (3) is separately coated with a detection line (5) composed of ractopamine-carrier protein conjugates and a control line (6) composed of rabbit anti-mouse IgG. The test strip of the present invention has advantages of high sensitivity and specificity, a simple operation, rapid detection and so on.

Description

technical field [0001] The invention belongs to the technical field of rapid immunochemical detection of veterinary drug residues, and specifically relates to an immunochromatographic reaction using colloidal gold markers for color development, and an immunocolloidal gold detection test paper for rapidly detecting ractopamine residues in pig urine. Background technique [0002] Ractopamine (Ractopamine) is a special kind of "Clenbuterol". Its efficiency is very high, adding less than 20g to one ton of feed can increase the lean meat of pigs in the final growth stage by 24% and reduce fat by 34%. This substance hardly accumulates in the body. Animal experiments and small-scale human experiments have found that when the intake per kilogram of body weight is below 67ug, there is no obvious damage to the human body. [0003] The allowable value of ractopamine residues established by the FDA is 50ppb (one part per billion, 50ppb is equivalent to 50 micrograms per kilogram) in po...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/558G01N33/566
Inventor 刘汉石
Owner DALIAN PRACTICAL BIOTECH
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