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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, high false negative rate and false positive rate, they are gradually replaced by high sensitivity, high accuracy and short detection time. short chemiluminescent immunoassay

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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Effect test

Embodiment 1

[0029] Example 1: Preparation of Specific Components of Prednisone Magnetic Immunochemiluminescence Detection Kit

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

[0031] Take 1.0 g of prednisolone, add methanol to dissolve, add 0.3 g of sodium hydride, stir, mix well, add 0.49 g of caproic hydrazide, stir, react at 70 ° C for 4 hours, stop the reaction, rotary steam, concentrate, add water, ethyl acetate Ester extraction, drying with anhydrous sodium sulfate and evaporating to dryness, putting on a silica gel column, eluting and separating with petroleum ether / ethyl acetate (V / V, 5 / 1) to obtain 1.2 g of hexanoylprednisone hapten product, yield 89% .

[0032] 2. Preparation of enzyme-labeled antigen

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

Embodiment 2

[0052] Example 2: Construction of Prednisone Magnetic Immunochemiluminescence Detection Kit

[0053] Construct the Prednisone Magnetic Immunochemiluminescent Detection Kit to include the following components:

[0054] (1) horseradish peroxidase-labeled prednisone hapten;

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

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

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

[0058] (5) The concentrations of prednisone series standard solutions are 0 μg / L, 0.1 μg / L, 0.3 μg / L, 0.9 μg / L, 2.7 μg / L, 8.1 μg / L;

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

Embodiment 3

[0062] Embodiment 3: the detection of residual amount of prednisone in the sample

[0063] 1. Sample pretreatment

[0064] Take 200 μL of milk sample, add 800 μL of reconstituted solution, oscillate with an oscillator for 2 min, and mix well; take 50 μL for analysis.

[0065] 2. Detection with kit

[0066] Dilute the enzyme-labeled antigen and the enzyme-labeled antigen diluent at a volume ratio of 1:10 to 1:20, and put it into the enzyme-labeled antigen working solution container of the chemiluminescent detector; dilute the magnetic-labeled antibody and the magnetic-labeled antibody diluent at 1:1 : Dilute at a volume ratio of 10 to 1:20, and put it into the magnetic label antibody working solution container of the chemiluminescence detector; set the position on the sample rack for each sample / standard product, 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, and the chemiluminescence detect...

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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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