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

An enzyme-linked immunosorbent reagent, tylosin technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of cumbersome processing and measurement operations, unsuitable for large-scale sample screening, sensitivity and specificity limitations, etc. The inspection method is convenient and easy to implement, shortens the inspection time, and has the effect of simple pretreatment process.

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1. Preparation and detection method of an enzyme-linked immunosorbent assay kit using a conjugate of a tylosin hapten and a carrier protein as a coating source

[0035]The ELISA kit using the conjugate of tylosin hapten and carrier protein as the coating source includes:

[0036] (1) A microtiter plate coated with a conjugate of tylosin and a carrier protein;

[0037] (2) Alkaline phosphatase-labeled goat anti-mouse anti-antibody working solution: Dilute the alkaline phosphatase-labeled goat anti-mouse anti-antibody to a protein concentration of 0.1-1 μg / L with enzyme-labeled antibody diluent. The alkaline phosphatase-labeled anti-antibody, the enzyme-labeled antibody diluent is a solution containing 0.1‰ (mass concentration) glycerin and 1% thimerosal preservative. 12ml / bottle, 1 bottle.

[0038] (3) Tylosin standard solution: 6 bottles of tylosin series standard solution, 0μg / L, 1.5μg / L, 4.5μg / L, 13.5μg / L, 40.5μg / L, 121.5μg / L, 1~3ml / bottle. The tylosin dru...

Embodiment 2

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

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

[0084] (1) Enzyme plate coated with tylosin anti-antibody;

[0085] (2) Alkaline phosphatase-labeled tylosin hapten working solution: Dilute the alkaline phosphatase-labeled tylosin hapten to a protein concentration of 0.1-1 μg / L with enzyme-labeled hapten diluent . The enzyme-labeled hapten diluent is a solution containing 50% glycerin and 1% thimerosal preservative. 12ml / bottle, 1 bottle.

[0086] (3) Tylosin standard solution: 6 bottles of tylosin series standard solution, 0μg / L, 1.5μg / L, 4.5μg / L, 13.5μg / L, 40.5μg / L, 121.5μg / L, 1~3ml / bottle. The tylosin drug dilution used is a phosphate buffer solution containing 5‰ (mass concentration) N,N'-dimethylformamide (DMF) and 1% ovalbumin (OVA).

[0087] (4) Chromogenic agent: 4-nitrophenol phosphate buffer. Both are 8ml / bottle.

[0088] (5) Tylosin rabbit ...

Embodiment 3

[0111] Embodiment 3, test kit precision, accuracy and shelf life test

[0112] 1. Kit precision test

[0113] (1) Precision test of standard product

[0114] Three batches of the kits prepared in Example 1 and Example 2 were taken respectively for precision experiments, and 10 kits were extracted 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 13.5 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 4.2% to 10.2%.

[0115] 1

2

3

4

5

6

7

8

9

10

CV%

01 batch

6.1

7.2

5.4

8.2

5.3

7.2

6.4

8.3

9.4

5.4

03 batches

5.4

6.1

7.2

8.3

4.2...

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PUM

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Abstract

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

Description

technical field [0001] The invention relates to a method for detecting tylosin and a special ELISA kit. Background technique [0002] The trade name of Halofuginone is Stenorol. It is a 0.6% premix of tylosin hydrobromide. It has special effects and has no cross-resistance with other coccidiostats, so it is often added to feed to control animal diseases caused by coccidia. It is a drug additive widely used in feed at present. However, tylosin is highly toxic to aquatic animals (such as fish and shrimp), and can harm humans through the food chain. If a person eats poultry tissue with a high residual concentration of tylosin, he or she will experience physical discomfort and other poisoning phenomena. 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 tylosin in chicken liver tissue is 130 μg / kg. [0003] At present, the methods common...

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张素霞吴小平汪善良
Owner CHINA AGRI UNIV
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