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Method for detecting salinomycin and spcial enzyme-linked immune reagent kit thereof

An enzyme-linked immunosorbent reagent, salinomycin technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of sensitivity and specificity limitations, cumbersome processing and measurement operations, and is not suitable for screening a large number of samples. The processing process is simple, the inspection method is convenient and easy, and the structure is simple

Inactive Publication Date: 2006-08-02
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the microbial detection method is economical and easy to operate, its sensitivity and specificity are limited when there are other microbial inhibitors in the sample; High sensitivity, but cumbersome sample pretreatment and determination operations, high cost, not suitable for screening a large number of samples, can be used as a confirmatory analysis of residues

Method used

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  • Method for detecting salinomycin and spcial enzyme-linked immune reagent kit thereof
  • Method for detecting salinomycin and spcial enzyme-linked immune reagent kit thereof
  • Method for detecting salinomycin and spcial enzyme-linked immune reagent kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1. Preparation and detection method of an enzyme-linked immunosorbent assay kit using a conjugate of a salinomycin hapten and a carrier protein as a coating source

[0033] The enzyme-linked immunosorbent assay kit using the conjugate of salinomycin hapten and carrier protein as the coating source includes:

[0034] (1) ELISA plates coated with salinomycin and carrier protein conjugates;

[0035] (2) Alkaline phosphatase-labeled goat anti-mouse anti-antibody working solution: Dilute the alkaline phosphatase-labeled goat anti-mouse anti-antibody with antibody diluent to a protein concentration of 0.1 μg / L, 12ml / bottle, 1 bottle. The enzyme marker diluent has a pH value of 8.2, contains 1.2 μg / L antibody protein, and contains 0.05 mol / L phosphate buffer with 5% methanol.

[0036] (3) Salinomycin standard solution: use diluent to dilute salinomycin into 6 bottles of series standard solution, 0μg / L, 1μg / L, 3μg / L, 9μg / L, 27μg / L, 81μg / L, 3ml / bottle. The salinomyci...

Embodiment 2

[0091] Example 2. ELISA kit using goat anti-rabbit anti-antibody as coating source and its preparation method

[0092] The ELISA kit with goat anti-rabbit anti-antibody as the coating source includes:

[0093] (1) Enzyme plate coated with salinomycin anti-antibody;

[0094] (2) Alkaline phosphatase-labeled salinomycin hapten working solution: dilute the alkaline phosphatase-labeled salinomycin hapten to 0.1 mol / L with double distilled water, 12ml / bottle, 1 bottle.

[0095] Salinomycin standard solution: use diluent to dilute salinomycin into 6 bottles of series standard solution, 0μg / L, 1μg / L, 3μg / L, 9μg / L, 27μg / L, 81μg / L, 3ml / bottle . The salinomycin drug diluent used is to contain 5‰N, N'-dimethylformamide (DMF), 1% bovine bovine serum (BSA) 0.05mol / L phosphate buffered saline

[0096] (4) The chromogenic solution is 4-nitrophenol phosphate buffer solution, 8ml / bottle, 1 bottle.

[0097] (5) Salinomycin rabbit polyclonal antibody working solution: dilute salinomycin rabb...

Embodiment 3

[0117] Embodiment 3, test kit precision, accuracy and shelf life test

[0118] 1. Kit precision test

[0119] (1) Precision test of standard product

[0120] 10 kits were drawn from each of the three batches of kits, and 20 microwells were drawn from the enzyme-linked plate of each kit, and the absorbance value (OD value) of the 9 μg / L standard solution was measured to calculate the coefficient of variation. The results are shown in Table 1, indicating that the coefficient of variation ranges from 4.8% to 16.2%, meeting the requirement that the precision is less than or equal to 20%.

[0121] 1

2

3

4

5

6

7

8

9

10

CV%

01 batch

6.2

7.4

9.5

10.5

16.2

4.9

8.2

9.2

7.5

6.4

03 batches

11.5

8.5

7.4

9.5

16.2

7.6

8.4

7.5

9.2

4.8

06 batches

8.5

6.4

7.2

9....

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PUM

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Abstract

The present invention discloses a method for detecting salinomycin and its special-purpose ELIA kit. Said kit includes salinomycin specific antibody, coating source and enzyme label. The described coating source is conjugate of salinomycin semiantigen and carrier protein or antiantibody, the described enzyme label is enzyme-labelled antiantibody or enzyme-labelled salinomycin semiantigen. When the described coating source is the conjugate of salinomycin semiantigen and carrier protein, the described enzyme label is enzyme-labelled antiantibody, and when the described coating source is antiantibody, the described enzyme label is enzyme-labelled salinomycin semiantigen.

Description

technical field [0001] The invention relates to a method for detecting salinomycin and a special ELISA kit. Background technique [0002] Salinomycin Sodium Premix is ​​a polyether veterinary antibiotic, which has antibacterial effects on most Gram-positive bacteria and some molds, and can exert a good preventive effect on chicken coccidiosis when used alone. However, the use of salinomycin should not be excessive. Excessive use will accumulate in large quantities in animals, and will poison and kill the animals eaten, and endanger human health through the food chain. In December 2002, my country's Ministry of Agriculture announced No. 235 that the maximum residue of salinomycin in the muscle of animals eaten was 600 μg / kg, the maximum residue in skin and fat was 1200 μg / kg in the liver It is 1800μg / kg. Therefore, it is very important to detect the highest residue level of salinomycin in food of animal origin. [0003] At present, the methods commonly used in the detection...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/577G01N33/535
Inventor 沈建忠何方洋万宇平史为民冯才伟江海洋吴小平汪善良
Owner CHINA AGRI UNIV
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