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Method for assaying sulfaquinoxaline and special enzyme-linked immune reagent kit

An enzyme-linked immunosorbent reagent, sulfaquinoxaline technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of complex equipment, unsuitable for on-site monitoring and screening of a large number of samples, and achieve high accuracy and inspection methods Convenient and easy to carry

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

[0003] The chemical methods for detecting sulfaquinoxaline residues mainly include thin layer chromatography (TLC), gas chromatography (GC), high pressure liquid chromatography (HPLC), gas-mass spectrometry (GC-MS), liquid-mass spectrometry (HPLC / MS), capillary electrophoresis (CE), etc., due to the complex equipment and cumbersome process, it is not suitable for on-site monitoring and screening of a large number of samples

Method used

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  • Method for assaying sulfaquinoxaline and special enzyme-linked immune reagent kit
  • Method for assaying sulfaquinoxaline and special enzyme-linked immune reagent kit
  • Method for assaying sulfaquinoxaline and special enzyme-linked immune reagent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1. Preparation and detection method of an enzyme-linked immunosorbent assay kit using a conjugate of a sulfaquinoxaline hapten and a carrier protein as a coating source.

[0037] The ELISA kit using the conjugate of sulfaquinoxaline hapten and carrier protein as the coating source includes:

[0038] (1) a microtiter plate coated with sulfaquinoxaline and carrier protein conjugate;

[0039] (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 0.5 μg / L. The diluent used for the enzyme marker working solution contains 0.05% glycerin (which can prevent the enzyme marker from being placed in the -20°C environment from freezing, and can also maintain the biological activity of the enzyme marker for a long time), 1% thimerosal preservative (convenient save) solution.

[0040] (3) sulfaquinoxaline standard substance solution: with the pH valu...

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) ELISA plate coated with goat anti-rabbit anti-antibody;

[0094] (2) Alkaline phosphatase-labeled sulfaquinoxaline hapten working solution: dilute the alkaline phosphatase-labeled sulfaquinoxaline hapten to a protein concentration of 0.5 μg / L. The diluent used for the enzyme marker working solution contains 0.05% glycerin (which can prevent the enzyme marker from being placed in the -20°C environment from freezing, and can also maintain the biological activity of the enzyme marker for a long time), 1% thimerosal preservative (convenient save) solution.

[0095] (3) Sulfaquinoxaline standard solution: 6 bottles of sulfaquinoxaline series standard solutions, 0μg / L, 1μg / L, 3μg / L, 9μg / L, 27μg / L, 81μg / L, 3ml / bottle.

[0096] (4) Chromogenic solution: 4-nitrophenol phos...

Embodiment 3

[0118] Embodiment 3, test kit precision, accuracy and shelf life test

[0119] 1. Kit precision test

[0120] (1) Precision test of standard product

[0121] 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 9 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 5.1% to 11.9%.

[0122] 1

2

3

4

5

6

7

8

9

10

CV%

01 batch

6.5

7.4

9.1

10.8

5.1

8.4

7.5

6.4

6.2

10.9

03 batches

11.9

8.5

6.4

7.5

9.4...

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Abstract

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

Description

technical field [0001] The invention relates to a method for detecting sulfaquinoxaline and a special ELISA reagent kit thereof. Background technique [0002] Sulfaquinoxaline (Sulfaquinoxaline SQX) belongs to sulfonamide antibiotics and has anticoccidial effect. It is widely used in poultry industry. It is absorbed quickly after oral administration, but excreted slowly, and remains in tissues, organs and eggs for a long time. After human consumption, Side effects such as drug resistance to the human body. The World Health Organization (WHO) stipulates that the maximum residue limit (MRL) value of animal tissue and milk is 100 μg / kg. Japan and other countries stipulate that it should not be detected in broiler chickens. Therefore, in order to ensure product safety and smooth foreign trade, it is very necessary to strengthen the detection of sulfaquinoxaline residues in animal foods. [0003] The chemical methods for detecting sulfaquinoxaline residues mainly include thin-...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/577G01N33/535
Inventor 沈建忠何方洋万宇平史为民李建成冯才伟吴小平
Owner CHINA AGRI UNIV
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