Method for detecting tumor marker by biochip

Abstract

The invention discloses a method for detecting a tumor marker by a biological chip, and relates to the technical field of tumor detection. The kit has high throughput, can detect multiple tumor markers at the same time, and is suitable for large-scale population screening; the sensitivity is high, and tumor markers at an extremely low level can be detected; high specificity is achieved, non-specific binding is reduced, and the detection accuracy is improved; the product has high stability and can be stored and transported for a long time at normal temperature.

Description

Technical Field

[0001] The present invention relates to the technical field of tumor detection, and particularly relates to a method for detecting tumor markers using a biochip. Background Art

[0002] A biochip, also known as a microarray or DNA chip, is a high-density microarray biosensor that immobilizes a large number of biomolecules (such as DNA, RNA, proteins, etc.) on a tiny solid matrix (such as a glass slide, silicon wafer, or plastic film, etc.). It utilizes the specific recognition reaction between biomolecules to enable rapid, efficient, and parallel detection of a large number of biomolecules in a biological sample within a very small space. Biochip technology is widely applied in fields such as genomics, transcriptomics, proteomics, etc., and has important application values in medical diagnosis, drug development, etc.

[0003] Tumor markers are chemical substances that reflect the presence of tumors. They either do not exist in normal adult tissues but are only found in embryonic tissues, or their content in tumor tissues is much higher than that in normal tissues. The presence or quantitative change of them can indicate the nature of the tumor, so as to understand the histogenesis, cell differentiation, and cell function of the tumor, and to assist in the diagnosis, classification, prognosis judgment, and treatment guidance of tumors.

[0004] Limitations of traditional cancer diagnosis methods: Traditional cancer diagnosis methods such as imaging examinations like X-ray, CT, MRI, etc. and histopathological examinations can, to a certain extent, detect tumors, but their sensitivity and specificity are often not high, and it takes a relatively long time to obtain results. In addition, these methods also have certain radiation risks and damage to the patient's tissues. Therefore, we propose a method for detecting tumor markers using a biochip. Summary of the Invention

[0005] The purpose of the present invention is to provide a method for detecting tumor markers using a biochip to solve the problems mentioned in the above background art.

[0006] The present invention specifically adopts the following technical solutions to achieve the above purpose:

[0007] A method for detecting tumor markers using a biochip includes:

[0008] Step 1, preparing a biochip and making a corresponding chip as a carrier;

[0009] Step 2, sample processing, collecting a blood sample from a patient and separating serum therefrom;

[0010] Step 3, sample detection, adding the separated serum to the prepared biochip for detection;

[0011] Step 4, Generate a detection report, and generate a detection report according to the sample detection results.

[0012] Further, the preparation of the biochip includes the following steps:

[0013] Step 11, Select a solid-phase carrier, fabricate the solid-phase carrier, and the solid-phase carrier is selected from one of a metal sheet, a silicon wafer, or a glass sheet;

[0014] Step 12, Fix the antibody, and fix the antibody on the solid-phase carrier by physical adsorption.

[0015] Further, the physical adsorption methods are: electrostatic interaction, hydrophobic interaction, and polar interaction.

[0016] Further, in the sample processing, serum separation is performed by centrifugation, which includes the following steps:

[0017] Step 21, Collection, collect whole blood in a blood collection tube without anticoagulant or with a coagulant promoter;

[0018] Step 22, Coagulation, let the sample coagulate naturally at room temperature;

[0019] Step 23, Centrifugation, centrifuge at 1500 g for 15 minutes at a temperature of 4 °C;

[0020] Step 24, Obtain serum, the supernatant is the serum;

[0021] Step 25, Sub-packaging, suck and sub-package into a clean cryopreservation tube with a pipette, and make marks;

[0022] Step 26, Quick-freezing, after the serum is sub-packaged into the cryopreservation tube, it should be immediately placed vertically in a storage device;

[0023] Step 27, Storage, transfer and store after quick-freezing.

[0024] Further, the coagulation time is 30 - 60 minutes, and it cannot be shaken or vibrated during this period.

[0025] Further, the storage device in the quick-freezing is liquid nitrogen, and the time is 30 seconds.

[0026] Further, the storage environment is liquid nitrogen or a -80 °C refrigerator.

[0027] Further, the sample detection includes the following steps:

[0028] Step 31, Bind the antibody, bind the tumor markers in the serum separated from the sample to the antibody fixed on the biochip;

[0029] Step 32: Generate a signal. By adding a specific monoclonal detection antibody against the anti-marker, generate an optical signal or other signals.

[0030] Step 33: Detect the signal. Use a biochip scanner to detect and analyze the generated signal.

[0031] Furthermore, the generation of the test report includes the following steps:

[0032] Step 41: Quantitative analysis. Perform quantitative detection of the tumor marker according to the signal intensity.

[0033] Step 42: Result reporting. Report the test results to the clinician for diagnosis and adjuvant treatment.

[0034] The beneficial effects of the present invention are as follows:

[0035] The present invention has high throughput, can detect multiple tumor markers simultaneously, and is suitable for large-scale population screening; it has high sensitivity and can detect extremely low levels of tumor markers; it has high specificity, reduces non-specific binding, and improves the accuracy of detection; it has high stability and can be stored and transported for a long time at room temperature. Detailed implementation manners

[0036] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be described clearly and completely below.

[0037] The present invention provides a method for detecting tumor markers using a biochip, including:

[0038] Step 1: Prepare a biochip. Fabricate a corresponding chip as a carrier.

[0039] Step 2: Sample processing. Collect the blood sample of the patient and isolate the serum therefrom.

[0040] Step 3: Sample detection. Add the serum after separation to the prepared biochip for detection.

[0041] Step 4: Generate a test report. Generate a test report according to the sample detection results.

[0042] In this embodiment, preferably, the preparation of the biochip includes the following steps:

[0043] Step 11: Select a solid-phase carrier. Fabricate a solid-phase carrier, and the solid-phase carrier is selected from one of a metal sheet, a silicon wafer, or a glass sheet; the metal sheet can meet the requirements of high-throughput screening, the silicon wafer is suitable for applications with high integration and precise control, and the glass sheet is suitable for various synthesis methods and has relatively simple pretreatment. In summary, the metal sheet, the silicon wafer, and the glass sheet can all be used as carriers, and the use effect is good.

[0044] Step 12: Fix the antibody. The antibody is fixed on the solid-phase carrier by physical adsorption.

[0045] In this embodiment, preferably, the physical adsorption methods are: electrostatic interaction, hydrophobic interaction, and polar interaction.

[0046] In this embodiment, preferably, centrifugation is used for serum separation in sample processing, which includes the following steps:

[0047] Step 21: Collection. Collect whole blood in a blood collection tube without anticoagulant or containing a clot activator.

[0048] Step 22: Coagulation. Let the sample coagulate naturally at room temperature.

[0049] Step 23: Centrifugation. Centrifuge at 1500 g for 15 minutes at 4 °C.

[0050] Step 24: Obtain serum. The supernatant is the serum.

[0051] Step 25: Aliquoting. Pipette and aliquot into clean cryotubes and make marks.

[0052] Step 26: Quick freezing. After the serum is aliquoted into cryotubes, it should be immediately placed vertically in a storage device.

[0053] Step 27: Storage. Transport and store after quick freezing.

[0054] In this embodiment, preferably, the coagulation time is 30 - 60 minutes, and it cannot be shaken or vibrated during this period.

[0055] In this embodiment, preferably, liquid nitrogen is used as the storage device for quick freezing, and the time is 30 seconds.

[0056] In this embodiment, preferably, the storage environment is liquid nitrogen or an -80 °C refrigerator.

[0057] In this embodiment, preferably, sample detection includes the following steps:

[0058] Step 31: Bind the antibody. Bind the tumor markers in the separated serum of the sample to the antibody fixed on the biochip.

[0059] Step 32: Generate a signal. Generate an optical signal or other signals by adding a specific monoclonal detection antibody against the marker.

[0060] Step 33: Detect the signal. Detect and analyze the generated signal using a biochip scanner.

[0061] In this embodiment, preferably, generating a test report includes the following steps:

[0062] Step 41, Quantitative analysis: quantitatively detect tumor markers according to the signal intensity;

[0063] Step 42, Result reporting: report the test results to clinicians for diagnosis and adjuvant treatment.

[0064] The working principle and usage process of the present invention:

[0065] Step 1, Prepare a biochip: fabricate a corresponding chip as a carrier, which includes the following steps:

[0066] Step 11, Select a solid-phase carrier: fabricate a solid-phase carrier, and the solid-phase carrier is selected from one of metal sheets, silicon wafers, or glass sheets;

[0067] Step 12, Immobilize the antibody: immobilize the antibody on the solid-phase carrier by physical adsorption, wherein the physical adsorption methods are: electrostatic interaction, hydrophobic interaction, and polar interaction.

[0068] Step 2, Sample processing: collect the blood sample of the patient and isolate the serum therefrom, which includes the following steps:

[0069] Serum separation in sample processing is performed by centrifugation, which includes the following steps:

[0070] Step 21, Collection: collect whole blood in a blood collection tube without anticoagulant or with a coagulant promoter;

[0071] Step 22, Coagulation: let the sample coagulate naturally at room temperature for 30 - 60 minutes, and do not shake or vibrate during this period;

[0072] Step 23, Centrifugation: centrifuge at 1500g for 15 minutes at 4°C;

[0073] Step 24, Obtain serum: the supernatant is the serum;

[0074] Step 25, Sub-packaging: suck and sub-package into a clean cryopreservation tube with a pipette, and make marks;

[0075] Step 26, Quick freezing: immediately place the serum sub-packaged in the cryopreservation tube vertically in liquid nitrogen for 30 seconds;

[0076] Step 27, Storage: transfer and store in liquid nitrogen or a -80°C refrigerator after quick freezing.

[0077] Step 3, Sample detection: add the serum after separation to the biochip after preparation for detection, which includes the following steps:

[0078] Step 31: Combine the tumor markers in the separated serum of the sample with the antibodies immobilized on the biochip.

[0079] Step 32: Generate signals. By adding specific monoclonal detection antibodies against the markers, optical signals or other signals are generated.

[0080] Step 33: Detect the signals. Use a biochip scanner to detect and analyze the generated signals.

[0081] Step 4: Generate a test report. Generate a test report according to the test results of the sample, which includes the following steps:

[0082] Step 41: Quantitative analysis. Quantitatively detect the tumor markers according to the signal intensity.

[0083] Step 42: Result reporting. Report the test results to the clinician for diagnosis and adjuvant treatment.

[0084] The above description of the disclosed embodiments enables those skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for detecting tumor markers using a biochip, characterized in that, Including: Step 1: Prepare a biochip and fabricate a corresponding chip as a carrier. Step 2: Sample processing, collect the patient's blood sample and isolate the serum therefrom. Step 3: Sample detection, add the serum after separation to the biochip after preparation for detection. Step 4: Generate a test report, generate a test report based on the sample detection results.

2. The method for detecting tumor markers using a biochip according to claim 1, wherein The preparation of the biochip includes the following steps: Step 11: Select a solid-phase carrier, fabricate the solid-phase carrier, and the solid-phase carrier is selected from one of a metal sheet, a silicon wafer, or a glass sheet. Step 12: Immobilize the antibody, immobilize the antibody on the solid-phase carrier by physical adsorption.

3. The method for detecting tumor markers by a biochip according to claim 2, wherein, The physical adsorption method is: electrostatic interaction, hydrophobic interaction, polar interaction.

4. A method for detecting tumor markers using a biochip according to claim 1, wherein, The serum separation in the sample processing is selected to be separated by centrifugation, which includes the following steps: Step 21: Collection, collect whole blood in a blood collection tube without anticoagulant or containing a coagulant promoter. Step 22: Coagulation, let the sample coagulate naturally at room temperature. Step 23: Centrifugation, centrifuge at 1500 g for 15 minutes under the condition of 4 °C. Step 24: Obtain serum, the supernatant is the serum. Step 25: Sub-packaging, aspirate and sub-package it into a clean cryogenic storage tube with a pipette and make marks. Step 26: Quick-freezing, the serum should be immediately placed vertically in a storage device after being sub-packaged into the cryogenic storage tube. Step 27: Storage, transport and store after quick-freezing.

5. The method for detecting tumor markers using a biochip according to claim 4, characterized in that, The coagulation time is 30 - 60 minutes, and it cannot be shaken or vibrated during this period.

6. The method for detecting tumor markers using a biochip according to claim 4, characterized in that, The storage device in the quick-freezing is selected as liquid nitrogen, and the time is 30 seconds.

7. A method for detecting tumor markers using a biochip according to claim 4, characterized in that, The storage environment is liquid nitrogen or an -80 °C refrigerator.

8. The method for detecting tumor markers using a biochip according to claim 1, characterized in that, The sample detection includes the following steps: Step 31: Bind the antibody, bind the tumor markers in the serum separated from the sample to the antibody immobilized on the biochip. Step 32: Generate a signal, generate an optical signal or other signals by adding a specific monoclonal detection antibody against the marker. Step 33: Detect the signal, use a biochip scanner to detect and analyze the generated signal.

9. A method for detecting tumor markers using a biochip according to claim 1, characterized in that The generation of the test report includes the following steps: Step 41: Quantitative analysis, quantitatively detect the tumor markers according to the signal intensity. Step 42: Result reporting, report the test results to the clinician for diagnosis and adjuvant treatment.