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Magnetic granule competing method chemiluminescence immune analysis determination reagent kit for detecting hormone and preparing method thereof

A chemiluminescence immunology and chemiluminescence technology, which is applied in the field of immunoanalysis medicine, can solve the problems of being unable to be widely used in clinical diagnosis and scientific research work, and restricting popularization and use, so as to achieve the effects of ensuring sensitivity, improving sensitivity, and facilitating operation and production

Inactive Publication Date: 2009-03-04
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The existing foreign chemiluminescence immunoassay kit is a closed automatic chemiluminescence measurement system, which requires an expensive automatic chemiluminescence measuring instrument, which limits the promotion and use, and cannot be widely used in clinical diagnosis and scientific research.

Method used

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  • Magnetic granule competing method chemiluminescence immune analysis determination reagent kit for detecting hormone and preparing method thereof
  • Magnetic granule competing method chemiluminescence immune analysis determination reagent kit for detecting hormone and preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Preparation of Thyroxine (T4) Magnetic Particle Competition Chemiluminescent Immunoassay Quantitative Assay Kit of the Present Invention (Using Alkaline Phosphatase to Prepare Labels)

[0041] 1. Marker preparation

[0042] Alkaline phosphatase was used to label the T4 antigen using the glutaraldehyde method, and the specific labeling process was as follows:

[0043] Mix according to the ratio of alkaline phosphatase: T4 antigen = 1:2, add glutaraldehyde for labeling with a volume of 10% of the mixed solution, mix well and react at room temperature for 1-2 hours. The solution was transferred into a dialysis bag, dialyzed overnight with 50 mM PBS solution of pH 7.4, and the medium was changed twice in the middle. Transfer the solution into a glass bottle, add an equal amount of high-quality glycerin, mix well, and store it below -20°C.

[0044] 2. Preparation of T4 calibrator

[0045] Prepared with T4 pure product, divided into 6 bottles of 0, 10, 25, 50, 15...

Embodiment 2

[0080] Example 2 Preparation of Thyroxine (T4) Magnetic Particle Competition Chemiluminescence Immunoassay Quantitative Assay Kit of the present invention (Horseradish Peroxidase is used to prepare markers)

[0081] The horseradish peroxidase-labeled T4 antigen adopts the sodium periodate oxidation method, and the specific labeling process is as follows:

[0082] Weigh 5 mg of HRP and dissolve in 1 mL of double distilled water. Add 0.2 mL of freshly prepared 0.1M NaIO to the above solution 4 The solution was stirred at room temperature in the dark for 20 minutes. Add 20 μL of 0.2M carbonate buffer at pH 9.5 to raise the pH of the above-formylated HRP to 9.0-9.5, then immediately add 10 mg of T4 antigen in 1 mL of 0.01M carbonate buffer, and gently Stir for 2 hours. Put the above solution into a dialysis bag, and dialyze against 0.15 M pH7.4 PBS at 4° C. overnight. Centrifuge at 10,000rpm for 30 minutes to remove the precipitate. The supernatant is the enzyme conjugate. Add...

Embodiment 3

[0084] Example 3 Preparation of the testosterone (T) magnetic particle competition method chemiluminescent immunoassay quantitative assay kit of the present invention

[0085] Except that the T antibody was used to coat the magnetic particles, the enzyme labeled T antigen was used to prepare the enzyme marker, and the concentration of the calibrator was 0, 0.5, 1.0, 2.5, 10, 20 ng / mL, the same method as in Example 1 and 2 was used to prepare T Chemiluminescent Immunoassay Quantitative Assay Kit for Magnetic Particle Competition Method.

[0086] The prepared test kit is tested by conventional manufacturing and testing procedures in this area, and the results are shown in the following table 2:

[0087] Table 2

[0088] Test items Inspection standards test result accuracy The average recovery rate is 90.0-110.0% 106.5％ specificity Cross-reaction rate with its analogues ≤0.01% 0.003％ Precision CV(%) ≤15％(n＝10) 6％ sensitivity ≤0.20ng / mL...

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Abstract

The invention relates to the immunoassay medical field, particularly provides a magnetic particle competition method chemiluminescence immunoassay assay kit and a preparation method thereof used for detecting hormones. The kit according to the invention comprises: 1) a calibrator; 2) magnetic particles which are coated with streptavidin; 3) hormone antigens of enzyme markers; and 4) a chemiluminescence substrate. Further, the method for preparing the kit according to the invention has the following steps: 1) pure raw materials are used to prepare the calibrator; 2) the antigens are used to coat the magnetic particles; 3) the antigens of the enzyme markers are prepared; 4) the calibrator, the chemiluminescence substrate and the antigens of the enzyme markers are packaged in a separated way; and 5) a finished product is packaged. The kit has the advantages of convenience, rapidness, sensitivity, stability, and the like.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine, in particular, the invention provides a magnetic particle competition chemiluminescence immunoassay quantitative assay kit and a preparation method thereof. Background technique [0002] Due to the surface effect, quantum size effect and macroscopic quantum tunneling effect of nanomaterials, nanomaterials present characteristics that conventional materials do not have in many fields. "Nanomaterials" and "nanotechnology" have penetrated into various fields of daily life. . Therefore, nanomaterials have very broad application prospects. [0003] The introduction of nanotechnology into modern medicine forms drug-loaded nanoparticles. The size of nanoparticles is generally much smaller than that of cells in organisms, which provides a new research path for biological research, that is, the use of nanoparticles for cell separation and utilization. Nanoparticles can be made into special drugs o...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N21/76G01N33/74
Inventor 唐宝军胡国茂应希堂
Owner CHEMCLIN DIAGNOSTICS CO LTD
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