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Magnetic microparticle chemiluminescence enzyme immune analytic reagent kit for detecting saccharide antigen and its use method

A technology of magnetic microparticles and chemiluminescent enzymes, which is applied in the biological field and achieves the effects of convenient and easy testing method, convenient carrying and wide linear range.

Inactive Publication Date: 2008-12-17
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, hospitals diagnose early tumors, mainly relying on imaging and cytopathological diagnostic techniques. Generally, they can be diagnosed only after they have obvious space-occupying lesions and clinical symptoms, but this is not the earliest stage of canceration.

Method used

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  • Magnetic microparticle chemiluminescence enzyme immune analytic reagent kit for detecting saccharide antigen and its use method
  • Magnetic microparticle chemiluminescence enzyme immune analytic reagent kit for detecting saccharide antigen and its use method
  • Magnetic microparticle chemiluminescence enzyme immune analytic reagent kit for detecting saccharide antigen and its use method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1. Preparation and detection method of magnetic microparticle chemiluminescent enzyme immunoassay kit for detecting CA50.

[0032] Magnetic microparticle chemiluminescent enzyme immunoassay kit for detection of CA50 includes:

[0033] (1) Magnetic microparticles coated with anti-FITC antibody: particle size 2-3μm, use concentration 5-10mg / mL, 20mL / bottle, 1 bottle;

[0034] (2) FITC-labeled CA50 monoclonal antibody (capture antibody) and enzyme-labeled CA50 monoclonal antibody (paired antibody) are mixed as a labeling solution: FITC-labeled capture antibody working solution and enzyme-labeled pairing at a volume ratio of 1:1 Antibody working solution is mixed, 15mL / bottle, 1 bottle. Among them, the FITC-labeled antibody solution is diluted to 1.25 μg / mL with 20% calf serum, and the diluent used for the enzyme working solution of labeled alkaline phosphatase is a thimerosal preservative solution containing 0.05% glycerin and 0.2g / L (which can prevent release of ...

Embodiment 2

[0068] Example 2. Preparation and detection method of magnetic microparticle chemiluminescent enzyme immunoassay kit for detecting CA125.

[0069] Magnetic microparticle chemiluminescent enzyme immunoassay kit for detection of CA125 includes:

[0070] (1) Magnetic microparticles coated with anti-FITC antibody: particle size 2-3μm, use concentration 5-10mg / mL, 20mL / bottle, 1 bottle;

[0071] (2) FITC-labeled CA125 monoclonal antibody (capture antibody) and enzyme-labeled CA125 monoclonal antibody (paired antibody) are mixed as a labeling solution: FITC-labeled capture antibody working solution and enzyme-labeled pairing at a volume ratio of 1:2 Antibody working solution is mixed, 15mL / bottle, 1 bottle. Wherein the FITC-labeled antibody solution is diluted to 1.25 μg / mL with 20% calf serum, and the diluent used for the enzyme working solution of the labeled horseradish peroxidase is a thimerosal preservative solution containing 0.05% glycerol and 0.2g / L (can be Prevent the enz...

Embodiment 3

[0107] Embodiment 3, kit precision, accuracy and stability experiment

[0108] 1. Determination of kit precision

[0109] (1) Standard precision experiment

[0110] Three batches of the kits prepared in Example 1 and Example 2 were taken respectively for the precision test, and 10 kits were taken from each batch. Use the kit extracted in Example 1 to measure the 40 U / mL CA50 standard 5 times, and use the kit extracted in Example 2 to measure the 100 U / mL CA125 standard 5 times. 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 4.0% and 8.2%.

[0111] Table 1 CA50 standard repeatability experiment

[0112]

[0113] The measurement results of the three batches of kits in Example 2 are shown in Table 2, and the results show that the coefficient of variation is between 5.8% and 8.8%.

[0114] Table 2...

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Abstract

The invention relates to a magnetic corpuscule chemiluminescent enzyme immunoassay kit for detecting carbohydrate antigen and the application method thereof. The kit comprises FITC antibody-coated magnetic corpuscules; a marker solution prepared by mixing the FITC-marked carbohydrate antigen monoclonal antibody and the enzyme-marked carbohydrate antigen monoclonal antibody; a carbohydrate antigen standard sample solution; a concentrated washing solution; and a luminescent substrate solution, wherein carbohydrate antigen optionally adopts one of CA72-4, CA50, CA19-9, CA242, CA15-3, CA27-29 and CA125. The enzyme-marked antibody and the FITC-marked antibody are the monoclonal antibodies corresponding to the antigens. The FITC antibody coating the magnetic corpuscules adopts a polyclonal antibody or a monoclonal antibody. The marker solution is prepared by mixing an FITC-marked capture antibody working solution and an enzyme-marked antibody pair working solution by the volume ratio of 1:(1-3). Compared with the known kit for mensurating the carbohydrate antigen, the kit has the advantages of high flux, high sensitivity, wide linear range, rapidness, etc., and has a wide application prospect for the clinical inspection, etc.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a magnetic particle chemiluminescent enzyme immunoassay kit for detecting carbohydrate antigens and a use method thereof. Background technique [0002] Cancer poses a serious threat to human survival and health. At present, the key to tumor treatment lies in early detection and early treatment, which has become a consensus. However, at present, hospitals diagnose early-stage tumors mainly by imaging and cytopathological diagnostic techniques. Generally, they are diagnosed only after they have obvious space-occupying lesions and clinical symptoms, but this is not the earliest stage of canceration. 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. The discovery of more and more tumor markers has put forward higher requirements for the establishment of new detection technolo...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/546
Inventor 林金明王栩
Owner TSINGHUA UNIV
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