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

An enzyme-linked immunosorbent reagent, the technology of hemosanone, which is applied in the field of detection of hemosanone, can solve the problems of cumbersome processing and measurement operations, unsuitability for large-scale sample screening, sensitivity and specificity limitations, etc. The effect of simple, simple sample preparation procedures

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1. Preparation and detection method of an enzyme-linked immunosorbent assay kit using a conjugate of a hemosanone hapten and a carrier protein as a coating source

[0034] The enzyme-linked immunosorbent assay kit using the conjugate of fusanone hapten and carrier protein as the coating source includes:

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

[0036] (2) Alkaline phosphatase-labeled goat anti-rabbit anti-antibody working solution: dilute the alkaline phosphatase-labeled goat anti-rabbit anti-antibody with antibody diluent to a protein concentration of 5.0 μg / L, 12ml / bottle, 1 bottle. The antibody diluent is pH 8.2, containing 1.2 μg / L antibody protein, and 0.05 mol / L phosphate buffer containing 5% methanol.

[0037] (3) Hemosanone standard solution: 6 bottles of hemosanone series standard solution, 0μg / L, 2μg / L, 6μg / L, 18μg / L, 54μg / L, 162μg / L, 1ml / bottle. The drug diluent of fushanone used is a phosphate buffer...

Embodiment 2

[0090] Example 2. ELISA kit using goat anti-mouse anti-antibody as coating source and its preparation method

[0091]ELISA kits with goat anti-mouse anti-antibody as the coating source include:

[0092] (1) A microtiter plate coated with goat anti-mouse anti-antibody;

[0093] (2) Horseradish peroxidase-labeled hrysanthone working solution: dilute the horseradish peroxidase-labeled hrysanthone hapten to 0.1 mol / L with double distilled water.

[0094] (3) Hemosanone standard solution: 6 bottles of hemosanone series standard solutions, 0μg / L, 2μg / L, 6μg / L, 18μg / L, 54μg / L, 162μg / L, 1ml / bottle. The drug diluent of fushanone used is a phosphate buffer containing 5‰ N,N'-dimethylformamide (DMF), 1% bovine bovine serum (BSA), and 0.05mol / L.

[0095] (4) Chromogenic solution A is hydrogen peroxide, and chromogenic solution B is tetramethylbenzidine, 7ml / bottle.

[0096] (5) Hemosanone mouse monoclonal antibody working solution: Dilute the monoclonal antibody secreted by the monoclo...

Embodiment 3

[0123] Embodiment 3, test kit precision, accuracy and shelf life test

[0124] 1. Kit precision test

[0125] (1) Precision test of standard product

[0126] Three batches were taken from the kits in Example 2 for the precision experiment, and 10 kits were taken from each batch of kits, and 20 microwells were extracted from the enzyme-linked plate of each kit to measure the 18 μg / L standard. The absorbance value (OD value) of the product solution was calculated to calculate the coefficient of variation. The measurement results are shown in Table 1, and the results show that the coefficient of variation ranges from 5.3% to 10.6%.

[0127] 1

2

3

4

5

6

7

8

9

10

CV%

01 batch

8.5

9.2

10.3

6.5

9.8

7.3

6.5

5.3

8.2

9.6

03 batches

7.5

7.9

9.4

6.8

10.6

12.3

7.7

8.2

9.6

8.4

06 ...

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PUM

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Abstract

The present invention discloses a method for detecting dichrone and its special-purpose ELIA kit. Said kit includes dichrone specific antibody, coating source and enzyme label. The described coating source is conjugate of dichrone semiantigen and carrier protein or antiantibody; the described enzyme label is enzyme-labelled antiantibody or enzyme-labelled dichrone semiantigen, when the described coating source is the conjugate of dichrone 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 dichrone semiantigen. The described dichrone semiantigen is dichrone derivative which is formed by means of reaction of ketone group of dichrone and carboxymethyl hydroxide radical and has carboxylic group.

Description

technical field [0001] The invention relates to a method for detecting hemosanone and a special ELISA kit. Background technique [0002] The trade name of Halofuginone is Stenorol, which is a 0.6% premix of halofuginone hydrobromide. It is a mixture of halofuginone A, B and C. Other anticoccidial drugs have no cross-resistance, so they are often added to feed to control animal diseases caused by coccidia, and are currently widely used drug additives in feed. However, Changshanone is highly toxic to aquatic animals (such as fish and shrimp), and can harm humans through the food chain. If people eat poultry meat tissue with a high residual concentration of fushanone, they will experience symptoms such as physical discomfort and other poisoning. Announcement No. 235 of the Ministry of Agriculture of the People's Republic of China "Maximum Residue Limits of Veterinary Drugs in Animal Foods" stipulates that the maximum residue limit of susanone in chicken liver tissue is 130 μg...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/577G01N33/535
Inventor 沈建忠丁双阳史为民何方洋张素霞万宇平江海洋刘金凤
Owner CHINA AGRI UNIV
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