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Method for detecting Ivermectin and special enzyme-linked immune reagent kit thereof

An enzyme-linked immunosorbent reagent, ivermectin technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of cumbersome processing and measurement operations, sensitivity and specificity limitations, unsuitable sample screening, etc., to achieve the goal of testing The method is convenient and easy, the pretreatment process is simple, and the effect of high accuracy

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 Ivermectin and special enzyme-linked immune reagent kit thereof
  • Method for detecting Ivermectin and special enzyme-linked immune reagent kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 The preparation and detection method of the enzyme-linked immunoassay kit using the conjugate of ivermectin hapten and carrier protein as the coating source

[0034] The ELISA kit using the conjugate of ivermectin hapten and carrier protein as the coating source includes:

[0035] (1) A microtiter plate coated with a conjugate of ivermectin and a carrier protein;

[0036](2) Alkaline phosphatase-labeled goat anti-mouse anti-antibody working solution: Use a diluent (containing 0.1‰ (mass concentration) glycerol (to prevent the enzyme marker from freezing at -20°C and keep it for a long time) Biological activity of enzyme markers), 1% calf serum, 1% thimerosal preservative (easy to store) solution) Dilute the alkaline phosphatase-labeled goat anti-mouse anti-antibody to a protein concentration of 0.1-1 μg / L Enzyme-labeled anti-antibody working solution, 12ml / bottle, 1 bottle.

[0037] (3) Ivermectin standard solution: Dilute ivermectin with a diluent into 6 bo...

Embodiment 2

[0082] Example 2, ELISA kit using anti-antibody as coating source and its preparation method

[0083] ELISA kits with ivermectin anti-antibody as coating source include:

[0084] (1) ELISA plates coated with anti-antibodies;

[0085] (2) Alkaline phosphatase-labeled ivermectin hapten working solution: Dilute alkaline phosphatase-labeled ivermectin hapten to 0.1-1 μg / L enzyme-labeled anti-antibody working solution, 12ml / bottle, 1 bottle. The diluent used contains 0.1‰ (mass concentration) glycerol (which can prevent the enzyme markers from freezing at -20°C and maintain the biological activity of the enzyme markers for a long time), 1% thimerosal preservative (easy to store) ) solution.

[0086] 3) Ivermectin standard solution: Dilute ivermectin with a diluent into 6 bottles of serial standard solutions, 0μg / L, 0.5μg / L, 1.5μg / L, 4.5μg / L, 13.5μg / L, 40.5 μg / L, 1~3ml / bottle. The ivermectin drug diluent used is a phosphate buffer solution with a pH value of 8.3 and 0.02 mol / L...

Embodiment 3

[0109] Embodiment 3, test kit precision, accuracy and shelf life test

[0110] 1. Kit precision test

[0111] (1) Precision test of standard product

[0112] Three batches of the kits prepared in Example 1 and Example 2 were taken respectively for the precision experiment, and 10 kits were taken from each batch of kits, and 20 microwells were respectively extracted from the enzyme-linked plate of each kit. Measure the absorbance value (OD value) of the 4.5 μg / L standard solution, and calculate the coefficient of variation. The measurement results of the three batches of kits in Example 1 are shown in Table 1, and the results show that the coefficient of variation ranges from 4.3% to 10.4%.

[0113] 1

2

3

4

5

6

7

8

9

10

CV%

01 batch

6.4

7.1

10.4

4.5

8.2

9.5

8.6

4.1

5.3

7.2

03 batches

6.2

7.4

8.1

9.2

5.4 ...

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PUM

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Abstract

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

Description

technical field [0001] The invention relates to a method for detecting ivermectin and a special ELISA kit. Background technique [0002] Ivermectin (Ivermectin) is a macrolide antibiotic, which can make chloride ions gather through the cell membrane, thereby causing paralysis of various nematodes and arthropods. It has multiple uses in the field of veterinary medicine. Ivermectin Toxic to arthropods and nematodes. However, high doses of ivermectin can cause the host to quickly clear the inflammatory response to nematodes or arthropods in the blood, resulting in adverse reactions to treatment. In December 2002, my country's Ministry of Agriculture announced No. 235 that the maximum residue in the muscle of all food animals was 10 μg / kg, and the maximum residue in fat was 40 μg / kg. Therefore, it is very important to detect the residual amount of ivermectin in veterinary medicine. [0003] At present, the methods commonly used in the detection of ivermectin residues mainly i...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/577G01N33/535
Inventor 沈建忠史为民何继红何方洋丁双阳万宇平
Owner CHINA AGRI UNIV
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