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Enzyme linked immunosorbent assay kit for detecting sulfadiazine and detection method thereof

A technology of sulfadiazine enzyme linkage and sulfadiazine, which is applied in the direction of biological testing, measuring devices, material inspection products, etc., can solve the problems of lack of sensitivity and specificity of sulfadiazine, difficulty in detecting samples, unfavorable promotion and application, etc., and achieve pre-treatment Simple process, low pre-treatment requirements, high sensitivity effect

Inactive Publication Date: 2014-05-21
JIANGSU WISE SCI & TECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, methods for detecting sulfadiazine include microbial methods, physical and chemical methods, such as high-pressure liquid chromatography (HPLC), gas-liquid chromatography (GLC), mass spectrometry (MS ), Thin Layer Chromatography (TLC), etc. Microbiological methods are faster in detection speed, but they lack sensitivity and specificity for sulfonamides. Physicochemical methods have the characteristics of cumbersome pretreatment, expensive instruments and professional operations, etc., to detect a large number of samples More difficult, not conducive to large-scale promotion and application

Method used

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  • Enzyme linked immunosorbent assay kit for detecting sulfadiazine and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Detection of animal tissue (chicken, chicken liver, pork, pork liver, fish, shrimp) samples

[0045] Pre-processing:

[0046] 1. Weigh 2.0±0.05 g homogenized sample into a 50 mL centrifuge tube;

[0047] 2. Add 8 mL of tissue sample extraction solution (see Preparation 3), and vortex for 3-5 minutes;

[0048] 3. Centrifuge at 4000 r / min for 10 min;

[0049] 4. Take 1 mL of the upper liquid in another centrifuge tube and dry it under nitrogen flow at 50~60°C;

[0050] 5. Add 1 mL of n-hexane to the centrifuge tube, then add 1 mL of diluted sample complex solution and shake well;

[0051] 6. Centrifuge at 4000 r / min for 5 min;

[0052] 7. Remove the upper phase, and take 50 μL of the lower layer for analysis.

[0053] Determination:

[0054] 1. Take the required reagents out of the refrigerated environment, and place them at room temperature (20-25°C) to equilibrate for more than 30 minutes. Note that each liquid reagent must be shaken well before use;

...

Embodiment 2

[0060] Example 2 Detection of milk samples

[0061] Pre-processing:

[0062] 1. Centrifuge the milk at room temperature at 4000 r / min for 15 minutes to remove the upper layer of fat (skimmed milk can omit this step);

[0063] 2. Dilute the centrifuged milk 5 times with deionized water (1 part milk + 4 parts deionized water) and shake well;

[0064] 3. Take 50 μL for analysis;

[0065] Determination: The specific determination steps are the same as in Example 1.

Embodiment 3

[0066] Example 3 Detection of honey samples

[0067] Pre-processing:

[0068] 1. Weigh 1±0.05 g of honey, add 2 mL of honey sample buffer (see recipe 4), and vortex until the honey is completely dissolved;

[0069] 2. Add 4 mL of acetonitrile and shake well;

[0070] 3. Centrifuge at 4000 r / min for 10 minutes at room temperature;

[0071] 4. Take 1 mL of the upper organic phase and dry it under nitrogen flow at 50~60°C;

[0072] 5. Add 1 mL of diluted sample reconstitution solution to dissolve the dried residue;

[0073] 6. Take 50 μL for analysis;

[0074] Determination: The specific determination steps are the same as in Example 1.

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Abstract

The invention discloses an enzyme linked immunosorbent assay kit for detecting sulfadiazine and a detection method thereof. The enzyme linked immunosorbent assay kit for detecting sulfadiazine includes a sulfadiazine specific antibody, and a coating sulfadiazine-carrier protein conjugate and an enzyme-labeled second antibody, or a coating second antibody and an enzyme-labeled sulfadiazine. The enzyme linked immunosorbent assay kit for detecting sulfadiazine can simultaneously detect large quantities of samples; the main reagents are all provided in a working liquid form, and the detection method is simple and is easy to implement; and the enzyme linked immunosorbent assay kit has the characteristics of high specificity, high sensitivity, high precision, high accuracy and the like, and can play an important role in detection of the residual amount of sulfadiazine in animal foodstuffs and fodders.

Description

technical field [0001] The invention relates to a sulfadiazine ELISA detection kit, which belongs to the technical field of biological and food safety detection. . technical background [0002] Sulfadiazine, as a kind of sulfonamide drug, has stable properties, broad antibacterial spectrum, low toxicity, easy oral absorption and low price. It is one of the most widely used drugs in animal husbandry. However, due to its long action time and metabolism time in the body, it is easy to remain in the animal body, and it may accumulate in the human body through any means. Drug accumulation concentration exceeding a certain value is harmful to human body function, and long-term accumulation will lead to the generation of sulfa drug resistance, resulting in epidemic infection of drug-resistant bacteria, and has potential carcinogenicity. Therefore, the European Union has set the maximum allowable value for sulfa drugs in milk and meat, that is, the total amount of sulfa drugs shal...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/531G01N33/543
CPCG01N33/54366G01N33/94
Inventor 杜道林张怿韡
Owner JIANGSU WISE SCI & TECH DEV
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