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Detection kit of acetyl amantadine for predicting tumors

A technology of acetylamantadine and a detection kit, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems that are not conducive to widespread promotion and use, require expensive instruments, and cumbersome operations, so as to be easy to promote and use on a large scale, eliminate False positive judgment and effect of improving specificity

Inactive Publication Date: 2011-05-04
BIOSINO BIO TECH & SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is cumbersome to operate and requires expensive instruments, which is not conducive to widespread use
In addition, diseases such as potential tumors, inflammation in the body, and myocardial ischemia can all lead to an increase in the concentration of acetylamantadine in the human body. This method cannot distinguish which disease caused the increase in the concentration of acetylamantadine. If the subject There are diseases such as inflammation and myocardial ischemia in the body, and it will also be wrongly judged as having a tumor or a potential tumor

Method used

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  • Detection kit of acetyl amantadine for predicting tumors
  • Detection kit of acetyl amantadine for predicting tumors
  • Detection kit of acetyl amantadine for predicting tumors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Preparation of monoclonal antibody A and monoclonal antibody B against acetylamantadine

[0039] The preparation process of anti-acetylamantadine monoclonal antibody A and monoclonal antibody B is as follows:

[0040] 1. Immunization: Rabbits were immunized with acetylamantadine as an antigen.

[0041]2. Fusion: Take out the spleen of the immunized rabbit, grind it, and fuse the spleen cells with myeloma cells. The fused hybridoma cells have the ability of spleen cells to secrete antibodies and the ability of unlimited proliferation of myeloma cells.

[0042] 3. Screening and cloning: divide the fused hybridoma cells into six 96-well plates for culture, take the cell culture supernatant, and detect whether it contains anti-acetylamantadine monoclonal antibody by ELISA method. The cell culture wells containing the anti-acetylamantadine monoclonal antibody were taken to perform limiting dilution method to clone hybridoma cells, and a hybridoma cell line capable...

Embodiment 2

[0050] Example 2 Detection kit for acetylamantadine for predicting tumors

[0051] Preparation method of the kit:

[0052] 1. Coating: Inject 100 μl of coating solution (5 μg / ml carbonate buffer solution) into the microwell plate, overnight at 4°C. The dosage of monoclonal antibody A is 5 μg / ml, 50 μl / well.

[0053] 2. Sealing: Take out the coated microwell plate and let it rest at room temperature for 30 minutes. Wash the plate once with a plate washer, pat dry the remaining water, inject 150 μl of blocking solution (2 g / ml fetal bovine serum) into the microwell plate, and overnight at 4°C.

[0054] 3. Drying: Take out the sealed microwell plate and let it rest at room temperature for 30 minutes. After discarding the blocking solution, pat dry the remaining water, and dry in a drying room until the residual liquid in the well evaporates. Then put it into an aluminum foil bag, seal it with a vacuum sealer, affix a label, and store it in a refrigerator at 4°C.

[0055] Sam...

Embodiment 3

[0068] Example 3 Combined application of immunological method for detection of acetylamantadine and biochemical method for detection of CRP and troponin

[0069] 1. Immunological detection method of acetylamantadine: double antibody sandwich method.

[0070] The specific method is as follows:

[0071] (1) Immobilizing the anti-acetylamantadine monoclonal antibody A on the carrier. Wherein, the anti-acetylamantadine monoclonal antibody A, the carrier is a microwell plate or magnetic beads.

[0072] (2) Add the sample to be tested (50 μl of the subject's urine) to combine the acetylamantadine with the monoclonal antibody A immobilized on the carrier.

[0073] (3) Wash the above-mentioned carrier with phosphate buffer saline, three times, 300 μl each time.

[0074] (4) Add 50 μl of anti-acetylamantadine monoclonal antibody B labeled with horseradish peroxidase or alkaline phosphatase to the washed carrier, wherein the anti-acetylamantadine monoclonal antibody B , so that the ...

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Abstract

The invention provides a detection kit of acetyl amantadine for predicting tumors. The kit comprises a carrier coated with an acetyl amantadine-resisting monoclonal antibody A and an acetyl amantadine-resisting monoclonal antibody B with a marker. The kit detects the acetyl amantadine in a sample on the basis of an immunological principle. The kit is used for detecting the acetyl amantadine for predicting the tumors, has the advantages of simplicity, fastness, high sensitivity and low cost and is easy to popularize and use on a large scale. In addition, the detection kit jointly applies an immunological method for detecting the acetyl amantadine and a biochemical method for detecting C-reactive protein (CRP) and troponin so as to eliminate false positive interference to tumor prediction due to diseases such as internal inflammation and myocardial ischemia and make the specificity of a method for early predicting cancers by detecting the acetyl amantadine higher.

Description

technical field [0001] The invention relates to a detection kit, in particular to a detection kit for acetylamantadine used for predicting tumors. Background technique [0002] Tumor markers refer to abnormally expressed active substances produced by tumor tissue or the body's response to tumor tissue, including hormones, enzymes, specific and non-specific glycoproteins or glycolipids, etc. They either do not exist in normal adult tissues but are only found in embryonic tissues, or their content in tumor tissues greatly exceeds that in normal tissues. Their existence or quantitative changes can indicate the nature of tumors, so as to understand the histogenesis and cell differentiation of tumors , cell function, to help tumor diagnosis, classification, prognosis judgment and treatment guidance. [0003] Polyamine is one of the growth factors of cells, and its abnormal increase can induce tumors. The content of polyamine in the urine of cancer patients is significantly highe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
Inventor 张志栋吴乐斌
Owner BIOSINO BIO TECH & SCI
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