Magnetic immunochemiluminescence detection kit for prednisone, and applications thereof

A chemiluminescence detection and chemiluminescence technology, applied in the field of immunological detection, can solve the problems of high false negative rate and false positive rate, low sensitivity, etc., and achieve the effect of short detection time, high sensitivity and specificity, and full automation.

Active Publication Date: 2018-08-03
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the most commonly used immunoassay methods include enzyme-linked immunoassay and colloidal gold immunochromatographic assay, but due to their low sensitivity, h

Method used

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  • Magnetic immunochemiluminescence detection kit for prednisone, and applications thereof
  • Magnetic immunochemiluminescence detection kit for prednisone, and applications thereof
  • Magnetic immunochemiluminescence detection kit for prednisone, and applications thereof

Examples

Experimental program
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Example Embodiment

[0029] Example 1: Preparation of specific components of a prednisone magnetic immunochemiluminescence detection kit

[0030] 1. Synthesis of prednisone hapten (see attached for synthetic route figure 1 )

[0031] Take 1.0g of cloprednisone, dissolve in methanol, add 0.3g of sodium hydride, stir and mix, add 0.49g of hexanoyl hydrazide, stir, react at 70°C for 4h, stop the reaction, rotary steam, concentrate, add water, ethyl acetate Ester extraction, dried with anhydrous sodium sulfate, evaporated to dryness, applied to a silica gel column, and eluted and separated with petroleum ether / ethyl acetate (V / V, 5 / 1) to obtain 1.2 g of hexanoyl prednisone hapten product with a yield of 89% .

[0032] 2. Preparation of enzyme-labeled antigen

[0033] Take 8mg of prednisone hexanoyl hapten, add 200μL of N,N-dimethylformamide to dissolve to obtain liquid A; take 100mg of horseradish peroxidase, add 6mL of water to dissolve, add 10mg of carbodiimide (EDC), Stir the reaction for 30 minutes to o...

Example Embodiment

[0052] Example 2: Construction of a prednisone magnetic immunochemiluminescence detection kit

[0053] Set up a prednisone magnetic immunochemiluminescence detection kit, which contains the following components:

[0054] (1) Horseradish peroxidase labeled prednisone hapten;

[0055] (2) Enzyme-labeled antigen diluent is pH 7.2~7.6, Na 3 PO 4 A buffer solution with a concentration of 0.01 mol / L and a NaCl concentration of 0.25 mol / L;

[0056] (3) Prednisone monoclonal antibody labeled with immunomagnetic beads;

[0057] (4) The diluent of the magnetic label antibody is pH 7.2~7.6, Na 3 PO 4 A buffer solution with a concentration of 0.01 mol / L and a NaCl concentration of 0.25 mol / L;

[0058] (5) The concentration of prednisone series standard solution is 0μg / L, 0.1μg / L, 0.3μg / L, 0.9μg / L, 2.7μg / L, 8.1μg / L;

[0059] (6) The chemiluminescence substrate A solution is a tris solution containing luminol and p-cresol, and the chemiluminescence substrate B solution is a solution containing citric a...

Example Embodiment

[0062] Example 3: Detection of residual prednisone in the sample

[0063] 1. Sample pretreatment

[0064] Take 200μL of milk sample, add 800μL of recombination solution, shake for 2min with a shaker, and mix; take 50μL for analysis.

[0065] 2. Detect with the kit

[0066] Dilute the enzyme-labeled antigen and the enzyme-labeled antigen diluent according to the volume ratio of 1:10~1:20, and put them into the chemiluminescence detector enzyme-labeled antigen working solution container; dilute the magnetic-labeled antibody and the magnetic-labeled antibody diluent according to 1 : Dilute with a volume ratio of 10~1:20, and put it into a chemiluminescence detector magnetic-labeled antibody working solution container; set the position on the sample rack for each sample / standard, enter the sample information and the name of the test item to be tested ; Put the sample tube / standard tube into the set sample rack, the chemiluminescence detector draws 50μL enzyme-labeled antigen working solu...

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Abstract

The invention provides a magnetic immunochemiluminescence detection kit for prednisone. The magnetic immunochemiluminescence detection kit comprises an enzyme labeled antigen, an enzyme labeled antigen dilution solution, a magnetically labeled antibody, a magnetically labeled antibody dilution solution, prednisone series standard solutions, a chemiluminescent substrate solution A, a chemiluminescent substrate solution B, a redissolution solution and a washing solution, wherein the enzyme labeled antigen is horseradish peroxidase-labeled prednisone hapten, and the magnetically labeled antibodyis immunomagnetic bead labeled prednisone monoclonal antibody. The invention further discloses a method for detecting the residual amount of prednisone in milk by using a chemiluminescence detector supporting the kit. According to the present invention, the method has advantages of high sensitivity, high specificity and short detection time in the detection of prednisone.

Description

technical field [0001] The invention relates to a chemiluminescence detection kit and a detection method thereof, in particular to a magnetic immunochemiluminescence detection kit for detecting the residual amount of prednisone in milk, belonging to the field of immunological detection. Background technique [0002] Prednisone belongs to glucocorticoid, which has a strong regulatory effect in the process of biosynthesis and metabolism, and also has the functions of suppressing immune response, anti-inflammation, anti-drug, and anti-shock. In livestock production, it is widely used because of its significant effect on improving the feed conversion rate, thereby promoting the growth and reproduction of livestock and aquatic organisms. However, in recent years, it has been abused in the process of animal husbandry due to reasons such as producers' unclear understanding of it, lax application of principles, and weak market supervision. Studies have found that drug residues caus...

Claims

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Application Information

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IPC IPC(8): G01N33/74
CPCG01N33/743G01N2430/00
Inventor 何方洋万宇平朱亮亮贾芳芳杜美红张颖杨春艳冯才伟
Owner BEIJING KWINBON BIOTECH
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