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Tumor-associated antigen 153 magnetic microparticle chemiluminescence immune assay determination kit and method for making same

A tumor-associated antigen and chemiluminescence technology, which is applied in the field of immunoanalysis medicine, can solve the problems of low sensitivity, short storage time of reagents, and low sensitivity

Inactive Publication Date: 2009-04-01
北京科美东雅生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Because radioimmunoassay technology uses radioactive elements as markers, it has certain pollution to the environment, and has disadvantages such as high instrument cost, low sensitivity, complicated operation, unstable measurement results, and short storage time of reagents.
Enzyme immunoassay has low sensitivity and many influencing factors, which can easily cause false negatives and false positives

Method used

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  • Tumor-associated antigen 153 magnetic microparticle chemiluminescence immune assay determination kit and method for making same
  • Tumor-associated antigen 153 magnetic microparticle chemiluminescence immune assay determination kit and method for making same
  • Tumor-associated antigen 153 magnetic microparticle chemiluminescence immune assay determination kit and method for making same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Preparation of Tumor-associated Antigen 153 Magnetic Particle Chemiluminescence Immunoassay Assay Kit of the present invention

[0042] 1. Preparation of Tumor-Associated Antigen 153 Calibrator

[0043] Dilute the pure product of tumor-associated antigen 153 with horse serum into calibrator, the concentrations are 0U / mL, 5U / mL, 15U / mL, 50U / mL, 150U / mL, 500U / mL, 6 bottles in total.

[0044] 2. Preparation of a mixture of tumor-associated antigen-153 monoclonal antibody-coated magnetic particles and biotin-labeled tumor-associated antigen-153 monoclonal antibody

[0045] 1. Preparation of magnetic particles coated with tumor-associated antigen 153 monoclonal antibody

[0046] Activate the magnetic particles with a particle size of 1-2 μm with glutaraldehyde, stir at room temperature, mix for 3 hours, apply a magnetic field, let stand for 20-25 minutes, pour out the supernatant, and use 0.01mol / L with a pH value of 7.4 Wash three times with phosphate buffer, an...

Embodiment 2

[0069] Embodiment 2 The usage method of the kit of the present invention

[0070] 1. Sample pretreatment

[0071] Take human fasting morning serum samples, centrifuge at 3000rpm for 5min, and take 50μL of supernatant for analysis.

[0072] 2. Detection method

[0073] Before using this kit for experiments, take out the mixture of antibody-coated magnetic particles and biotin-labeled antibodies, calibrator / sample to be tested, and enzyme-labeled avidin and place them at room temperature for 15-30 minutes to allow them to balance to Room temperature; after that, adjust the incubator or water bath to 37°C; then, prepare a suitable micro-sampler and corresponding tips and check whether the chemiluminescence instrument is working normally.

[0074] The specific operation steps of using this kit to carry out the experiment according to the method of embodiment 1 are as follows:

[0075] After numbering the round-bottom polystyrene test tubes, add 50 μL of serum samples or serial ...

Embodiment 3

[0076] The methodological examination of the kit of the present invention of embodiment 3

[0077] The test kit prepared in Example 1 is tested according to the conventional manufacturing and testing procedures in the art, and the results are as follows:

[0078] 1. Determination of kit precision

[0079] (1) Calibrator precision experiment

[0080] Three batches of the kits prepared in Example 1 were taken for the precision test, and 10 kits were taken from each batch. Measure the 50 U / mL CA153 calibrator 5 times with the kit extracted in Example 1. Calculate the coefficient of variation for the measured concentrations. 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 is between 3.5% and 8.1%.

[0081] Table 1 CA153 Calibration Repeatability Experiment

[0082]

[0083] (2) Sample repeatability experiment

[0084] Take two normal human serum, add CA153 calibrator to 50U / mL a...

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Abstract

The invention relates to the medical field of immunoassay, particularly provides a measuring kit of the chemiluminescence immunoassay of a CA153 magnetic particle. The kit of the invention comprises: 1) a CA153 working calibrator; 2) the mixed solution of the magnetic particle coated with a CA153 monoclonal antibody and the CA153 monoclonal antibody marked by biotin; 3) streptavidin marked by horse radish peroxidase; 4) a chemiluminescence substrate; and 5) a reaction tube. Further, the method for preparing the kit according to the invention comprises the following steps: 1) the working calibrator is prepared by CA153 pure products; 2) the mixed solution of the magnetic particle coated with the monoclonal antibody and the monoclonal antibody marked by the biotin is prepared; 3) the horse radish peroxidase is used for marking the streptavidin; 4) the chemiluminescence substrate is prepared; 5) the working calibrator, the mixed solution, enzyme markers, the chemiluminescence substrate and the reaction tube are sub-assembled; and 6) assembly is carried out to obtain a finished product. The kit of the invention has the advantages of simpleness and convenience, fastness, sensitivity and stability and the like.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine, in particular, the invention provides a tumor-associated antigen 153 magnetic particle chemiluminescence immunoassay assay kit and a preparation method thereof. The kit of the invention combines biotin-avidin immunomagnification technology, immunomagnetic particle technology and chemiluminescence immunoassay technology. Background technique [0002] Cancer has posed a serious threat to human survival and health since its birth. At present, the diagnosis of early tumors in hospitals mainly relies on imaging and cytopathological diagnostic techniques, and the diagnosis is usually made after there are obvious space-occupying lesions and clinical symptoms. With the early diagnosis of tumors entering the protein era, body fluid proteins represented by blood have become a hot spot in the field of tumor marker research, thus promoting the establishment and development of new detection technologie...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/574G01N21/76
Inventor 王栩林金明李振甲应希堂胡国茂
Owner 北京科美东雅生物技术有限公司
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