Male multi-tumor marker detection protein chip and kit thereof

Abstract

The invention discloses a male multi-tumor marker detection protein chip and a kit thereof. The chip comprises a substrate, protein markers distributed in an array type and point coatings of contrasts, wherein the substrate is a glass substrate or a film substrate; and the tumor markers and the point coatings of the contrasts are seven protein markers of AFP, CEA, NSE, CYFRA21-1, CA19-9, tPSA and SCC-ag, a positive contrast and a negative contrast which are uniformly distributed and latticed on the substrate. A reaction result of various indexes can be obtained only through one reaction by utilizing the protein chip, and the following tumors can be simultaneously screened: primary liver cancer, prostatic cancer, pancreatic cancer, lung cancer, esophageal cancer, gastric cancer and colorectal cancer. The invention is particularly suitable for the general examination of malignant tumors of male asymptomatic groups and high risk groups.

Description

technical field

[0001] The invention relates to a protein chip disease detection product, in particular to a male multi-tumor marker detection protein chip and a kit thereof. Background technique

[0002] Early diagnosis of malignant tumors can improve the prognosis and survival rate of patients. The clinical diagnosis of male malignant tumors (including primary liver cancer, prostate cancer, pancreatic cancer, lung cancer, esophageal cancer, gastric cancer, and colorectal cancer) generally includes imaging examinations (such as B-ultrasound, X-ray barium meal, X-ray, CT, MRI), serum tumor marker detection (such as tPSA, AFP, CA19-9, etc.), digital rectal examination, endoscopy (esophageal endoscopy, fiber optic endoscopy, Folding sigmoidoscopy, total colonoscopy) and biopsy. Although imaging and endoscopy can detect malignancies early, they cannot be used for physical examination screening in the general population because they are expensive or invasive. However, most mal...

Examples

Embodiment 1

[0028] Preparation of a protein chip for detection of multiple tumor markers in males

[0029] The main steps are: (1) Prepare the matrix nitrocellulose membrane; (2) Design the chip, determine the arrangement of the lattice and the spotting position; (3) Draw the antibody from the 384-well plate and spray it on the nitrocellulose (4) assemble a nitrocellulose membrane, and carry out sealing, drying, packaging, and preservation.

[0030] The preparation method of the above-mentioned protein chip for detecting multiple tumor marker antibodies in males specifically includes the following steps:

[0031] 1. Determination of antigen and antibody for tumor marker detection

[0032] Determination of detection index antigen:

[0033] The tumor marker CEA is composed of six peptides with high similarity, so the A1+B1 segment was selected for genetic engineering expression; PSA, NSE, SCC-ag and CYFRA21-1 are full-length genetic engineering expression; while AFP and CA19-9 are purifi...

Embodiment 2

[0041] Kit preparation

[0042] A kit for male multi-tumor marker detection protein chip, composed of male multi-tumor marker detection protein chip, standard product, quality control product, sample diluent, enzyme-labeled working solution, concentrated washing solution, and detection solution; each part For bottled packaging, use a paper template to fix in the box (such as figure 2 ), not limited to the placement method, it is used together with the test. Here's how the parts are prepared:

[0043]1. The male multi-tumor marker detection protein chip is a 6×8-well plate, with nitrocellulose membrane as the base, and the assembly method is shown in ZL032298900. After the completion of the appearance, see figure 1 .

[0044] 2. The standard product provides 6 bottles, each bottle is 0.25ml.

[0045] With 3% BSA and 0.05% preservatives (Proclin TM 300) of phosphate buffer (pH 7.4) configuration, wherein the final concentrations of AFP, CEA, CA19-9, tPSA, NSE, SCC-ag and...

Embodiment 3

[0063] Application and detection of protein chips and kits

[0064] The application of the protein chip for detecting multiple tumor marker antibodies in males according to the present invention mainly adopts the antibody-antigen-enzyme-labeled secondary antibody method to detect tumor marker antibodies in human blood, and the method is to add human serum to the surface of the chip , the antigens in the sample are bound to the corresponding antibodies immobilized on the chip, after drying, add the enzyme-labeled secondary antibody to react, wash away the unbound secondary antibody, scan and collect the signal 1 minute after adding the detection solution, and the sample point The intensity of the signal is proportional to the concentration of the antigen in the serum.

[0065] In the above-mentioned application of the protein chip kit for detection of multiple tumor marker antibodies in males, the specific methods of antigen-antibody reaction and detection are as follows:

[0...

technical field

[0001] The invention relates to a protein chip disease detection product, in particular to a male multi-tumor marker detection protein chip and a kit thereof. Background technique

[0002] Early diagnosis of malignant tumors can improve the prognosis and survival rate of patients. The clinical diagnosis of male malignant tumors (including primary liver cancer, prostate cancer, pancreatic cancer, lung cancer, esophageal cancer, gastric cancer, and colorectal cancer) generally includes imaging examinations (such as B-ultrasound, X-ray barium meal, X-ray, CT, MRI), serum tumor marker detection (such as tPSA, AFP, CA19-9, etc.), digital rectal examination, endoscopy (esophageal endoscopy, fiber optic endoscopy, Folding sigmoidoscopy, total colonoscopy) and biopsy. Although imaging and endoscopy can detect malignancies early, they cannot be used for physical examination screening in the general population because they are expensive or invasive. However, most mal...

Examples

Embodiment 1

[0028] Preparation of a protein chip for detection of multiple tumor markers in males

[0029] The main steps are: (1) Prepare the matrix nitrocellulose membrane; (2) Design the chip, determine the arrangement of the lattice and the spotting position; (3) Draw the antibody from the 384-well plate and spray it on the nitrocellulose (4) assemble a nitrocellulose membrane, and carry out sealing, drying, packaging, and preservation.

[0030] The preparation method of the above-mentioned protein chip for detecting multiple tumor marker antibodies in males specifically includes the following steps:

[0031] 1. Determination of antigen and antibody for tumor marker detection

[0032] Determination of detection index antigen:

[0033] The tumor marker CEA is composed of six peptides with high similarity, so the A1+B1 segment was selected for genetic engineering expression; PSA, NSE, SCC-ag and CYFRA21-1 are full-length genetic engineering expression; while AFP and CA19-9 are purifi...

Embodiment 2

[0041] Kit preparation

[0042] A kit for male multi-tumor marker detection protein chip, composed of male multi-tumor marker detection protein chip, standard product, quality control product, sample diluent, enzyme-labeled working solution, concentrated washing solution, and detection solution; each part For bottled packaging, use a paper template to fix in the box (such as figure 2 ), not limited to the placement method, it is used together with the test. Here's how the parts are prepared:

[0043]1. The male multi-tumor marker detection protein chip is a 6×8-well plate, with nitrocellulose membrane as the base, and the assembly method is shown in ZL032298900. After the completion of the appearance, see figure 1 .

[0044] 2. The standard product provides 6 bottles, each bottle is 0.25ml.

[0045] With 3% BSA and 0.05% preservatives (Proclin TM 300) of phosphate buffer (pH 7.4) configuration, wherein the final concentrations of AFP, CEA, CA19-9, tPSA, NSE, SCC-ag and...

Embodiment 3

[0063] Application and detection of protein chips and kits

[0064] The application of the protein chip for detecting multiple tumor marker antibodies in males according to the present invention mainly adopts the antibody-antigen-enzyme-labeled secondary antibody method to detect tumor marker antibodies in human blood, and the method is to add human serum to the surface of the chip , the antigens in the sample are bound to the corresponding antibodies immobilized on the chip, after drying, add the enzyme-labeled secondary antibody to react, wash away the unbound secondary antibody, scan and collect the signal 1 minute after adding the detection solution, and the sample point The intensity of the signal is proportional to the concentration of the antigen in the serum.

[0065] In the above-mentioned application of the protein chip kit for detection of multiple tumor marker antibodies in males, the specific methods of antigen-antibody reaction and detection are as follows:

[0...

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