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Tumour serum mark and its uses

A technology of serum markers and tumors, applied in the field of tumor serum markers and its application, can solve the problems that the sensitivity and specificity of tumor serum markers cannot meet clinical diagnosis

Inactive Publication Date: 2008-01-09
SHANDONG MEDICAL BIO TECH RES CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] As the sensitivity and specificity of existing tumor serum markers cannot meet the needs of clinical diagnosis, the present invention proposes a method based on the study of the expression and mechanism of peptidyl arginine deiminase 4 (PADI4) in tumor tissues. A tumor serum marker and its application in the preparation of clinical diagnostic reagents or kits for detecting tumors

Method used

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  • Tumour serum mark and its uses
  • Tumour serum mark and its uses
  • Tumour serum mark and its uses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1. Enzyme-linked immunosorbent assay (ELISA) measures the expression of PADI4 in serum

[0030] 1. Reagent Preparation

[0031] (1) 0.05M carbonate buffer (Carbonate-bicarbonate buffer) coating diluent, pH9.6:

[0032] Sodium carbonate (Na 2 CO 3 ) 1.59g, sodium bicarbonate (NaHCO 3 ) 2.93g, sodium azide (sodium azide) 0.2g, add water to 1000 ml, adjust the pH to 9.6.

[0033] (2) PBS buffer (pH7.4-7.6):

[0034] Sodium chloride (NaCl) 8g, anhydrous disodium hydrogen phosphate (NaCl) 2 HPO 4 ) 1.15g, potassium dihydrogen phosphate (KH 2 PO 4 ) 0.2g, potassium chloride (KCl) 0.2g, add water to 1 liter.

[0035] (3) Blocking solution:

[0036] Add 3 g of skimmed milk powder to 100 ml of PBS buffer (pH7.4) solution.

[0037] (4) PBS-Tween buffer (PBST) washing solution, pH7.4:

[0038] Add 0.5 ml of Tween-20 (Tween-20) to 1 liter of the above PBS buffer solution (pH7.4), add water to 1 liter, and adjust the pH to 7.4.

[0039] 2. The experiment was ca...

Embodiment 2

[0049] Embodiment 2: preparation ELISA (direct method) detects serum PADI4 kit

[0050] Serum PADI4 detection kit contains:

[0051] (1) ELISA plate (strip);

[0052] (2) 0.05M carbonate buffer sample coating diluent, pH9.6;

[0053] (3) Blocking solution: PBS buffer solution (pH7.4) containing 3% skimmed milk powder;

[0054] (4) Anti-PADI4 antibody, the antibody is routinely labeled with horseradish peroxidase;

[0055] (5) PBS antibody diluent: PBS buffer containing 2% BSA;

[0056] (6) Washing solution: PBS buffer containing 0.05% Tween-20 (PBST);

[0057] (7) Chromogen: 0.4% tetramethylbenzidine (TMB);

[0058] (8) Stop solution: 2% H 2 SO 4 .

[0059] (9) PADI4 recombinant protein or positive and negative control substances.

Embodiment 3

[0060] Embodiment 3: ELISA (direct method) detects the application of serum PADI4 kit

[0061] (1) Take the required amount of pre-coated plates (strips), dilute the serum to be tested 10-20 times with the sample diluent, add 100ul per well to the wells of the microplate, and place the microplate in a wet box at 4 degrees Celsius overnight.

[0062] (2) Discard the sample solution, add 200ul of PBST to each well, and then discard the PBST. Wash the plate 3 times with washing solution.

[0063] (3) Add 200ul of blocking solution to each well, and react the microplate plate in a humid box at room temperature for half an hour.

[0064] (4) Discard the blocking solution, and add 100ul of anti-PADI4 antibody to each well. The microtiter plate was reacted in a humid box at room temperature for 2 hours.

[0065] (5) Discard the primary antibody solution, and wash the microtiter plate three times with PBST.

[0066] (6) Add 100ul of chromogen to each well, and carry out chromogen...

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Abstract

The invention disclose a pulchritude of neoplasm sera, which made up of peptide metarginase; among of which tumor is that one of breast carcinoma, liver cancer, renal carcinoma, oophoroma, carcinoma of prostate and bladder carcinoma. The invention also disclose the application of pulchritude of neoplasm sera in the clinical diagnosis reagent of preparing detect tumor. Using the tumor clinical diagnosis reagent or kit which is provided by this invention and prepared pulchritude of neoplasm sera, it can detect a guideline and realize preparatory ensure if the tester suffer from malignancy, accomplish the work of health screening with low cost in the short detect time, and provide the reliable basis for the clinical diagnosis.

Description

technical field [0001] The present invention relates to a serum marker and its application, in particular to a tumor serum marker and its application. Background technique [0002] Tumor markers refer to tumor-associated antigens, active proteins, hormones, receptors, enzymes or isoenzymes directly synthesized and secreted by tumor cells, some of which can enter blood or body fluids through circulation. Tumor markers in blood and body fluids can be detected by biochemical reaction, radioimmunoassay, enzyme immunoassay, electrochemiluminescence and other methods. [0003] The discovery of tumor markers is of great significance for tumor diagnosis, treatment, monitoring recurrence, judging efficacy and anticancer drug design. At present, there are dozens of tumor markers commonly used in hospitals, such as AFP (alpha-fetoprotein), CEA (carcinoembryonic antigen), CA125 (carbohydrate antigen 125), CA199 (carbohydrate antigen 199), CA153 (carbohydrate antigen 153), CA50 (carboh...

Claims

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Application Information

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IPC IPC(8): G01N33/52G01N33/53
Inventor 常晓天朱有名韩金祥朱波
Owner SHANDONG MEDICAL BIO TECH RES CENT
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