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Application of peptidylarginine deiminase 2 (PAD2) to preparation of reagent for clinical diagnosis of tumors

A technology of peptidyl arginine deiminase and diagnostic reagents, which is applied in the field of application of peptidyl arginine deiminase 2 in the preparation of tumor clinical diagnostic reagents, and can solve problems such as distribution of different tissues

Active Publication Date: 2015-02-18
SHANDONG XINCHUANG BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although PAD2 (peptidyl arginine deiminase 2, Peptidylarginine deiminase 2, or PADI2) and PAD4 are both peptidyl arginine deiminases, they have different tissue distributions. Currently, there is no research on the role of PAD2 in tumors. Related reports on blood markers

Method used

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  • Application of peptidylarginine deiminase 2 (PAD2) to preparation of reagent for clinical diagnosis of tumors
  • Application of peptidylarginine deiminase 2 (PAD2) to preparation of reagent for clinical diagnosis of tumors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 PAD2 as a detection example of tumor blood markers

[0054] PAD2 was used as a tumor blood marker to detect the blood of tumor patients. The number of tumor patients and the types of tumors are shown in Table 1.

[0055] The detection method is:

[0056] (1) Dilute the blood sample to be tested 10 times with carbonate buffer (pH 9.6), add it to a blank microtiter plate, and incubate at 37°C for 2 hours;

[0057] (2) discard the liquid in the hole, and wash the plate three times with PBST washing solution (the PBST washing solution refers to the PBS solution plus Tween-20, the pH of the PBS solution is 7.4, and the concentration of Tween-20 is 0.1%, percent by volume);

[0058] (3) Add 200 μL / well of blocking solution (0.5% BSA, mass percent) to each well, and incubate at 37°C for 1 hour;

[0059] (4) Discard the liquid in the well, and wash the plate three times with PBST washing solution;

[0060] (5) Add 100 μL / well of PAD2 antibody dilution (1:4000 dilut...

Embodiment 2 PAD2 test example 2

[0074] Using PAD2 as a tumor blood marker, the patient's blood was detected, and the detection method was the same as in Example 1. At the same time, with CEA, CA199, F / PSA, CA125, PSA, CA242 in the prior art as tumor blood markers, the blood of tumor patients is detected (the detection method is a conventional method in the prior art; these prior art The blood sample detection data of the tumor blood markers in is the patient outpatient data of a certain hospital, and this data can be used as one of the basis for diagnosing whether the patient has a tumor; the present invention also uses the blood samples of some patients to detect PAD2). The positive rate of each tumor blood marker was compared, and the cross-comparison between PAD2 and each tumor marker.

[0075] Results: The number of patients (a total of 906 cases) and the number of positive cases are shown in Table 3. It can be seen from Table 3 that PAD2 can be used as a tumor blood marker with good specificity.

[00...

Embodiment 3 PAD2 test example 3

[0079] Using PAD2 as a blood marker, the blood of patients with hepatitis B, general inflammation (increased leukocytes, neutrophils, and decreased lymphocyte ratio), kidney disease (including nephrotic syndrome, renal failure, and nephritis) was detected. See Table 4 for details of the number and types of diseases.

[0080] The detection method is the same as in Example 1.

[0081] Result: The number of positive cases and the positive rate are shown in Table 4. As can be seen from Table 4, PAD2, as a blood marker, can also be used to detect hepatitis B, common inflammation (with leukocytes, neutrophils, symptoms of decreased lymphocyte ratio) ), kidney disease (including nephrotic syndrome, renal failure, nephritis) and other diseases, the test results can be used as one of the basis for clinical diagnosis.

[0082] Table 4

[0083] disease category

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Abstract

The invention discloses an application of peptidylarginine deiminase 2 (PAD2) to preparation of a clinically diagnostic reagent or detection object for diagnosing tumors. The tumors are selected from liver cancers, gastric cancers, lung cancers, breast cancers, esophageal cancers, ovarian cancers, uterine cancers and colorectal cancers. When PAD2 is specially applied, an antibody against PAD2 is prepared by a conventional method, and a qualitative or quantitative method and a matching kit for detecting PAD2 are established. The invention also discloses a blood diagnosis reagent for diagnosing tumors. The blood diagnosis reagent comprises the PAD2 antibody, a horseradish peroxidase-immunoglobulin G (HRP-IgG) antibody and conventional reagents in the blood diagnosis reagent, wherein the conventional reagents in the blood diagnosis reagent include a carbonate buffer solution, a phosphate buffer solution-Tween (PBST) lotion, a blocking solution, developing solutions and a stop solution. PAD2 is found for the first time to be capable of being applied as a tumor blood marker. Experiments prove that PAD2 has feasibility as the tumor marker.

Description

technical field [0001] The invention relates to the application of peptidyl arginine deiminase 2 as a tumor blood marker in the preparation of tumor clinical blood diagnosis reagents. Background technique [0002] Peptidyl arginine deiminase (Peptidylarginine deiminase, PAD or PADI) is an enzyme in human tissues. At present, a total of five PAD enzymes (ie, PAD1, 2, 3, 4 and 6) have been discovered. These enzymes are encoded by a cluster of genes located on the lp36 region of the human chromosome and have distinct tissue distributions. It can perform post-translational modification on some other tissue proteins in the presence of calcium ions. This enzyme can catalyze the amino group of arginine (arginine) in the polypeptide chain into a carbonyl group, thereby converting arginine into citrulline (citrulline). Citrulline is an unnatural amino acid. This process of converting arginine to citrulline in polypeptides catalyzed by PAD is called citrullination. After citrullin...

Claims

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Application Information

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IPC IPC(8): G01N33/577
CPCG01N33/577G01N33/574
Inventor 常晓天邢艳秋马芳杨冬霞
Owner SHANDONG XINCHUANG BIOLOGICAL TECH CO LTD
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