CN-121978338-A - Kit and detection method for detecting ovarian cancer

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to a kit and a detection method for detecting ovarian cancer. The kit comprises an ELISA plate and tumor-associated antigens coated on the ELISA plate, wherein the tumor-associated antigens consist of a combination of Sui1, NDE1 and PDLIM 1. The kit provided by the invention uses three antigens, namely Sui1, NDE1 and PDLIM1, as a combination for screening ovarian cancer for the first time, can specifically detect the content of the Sui1, NDE1 and PDLIM1 autoantibodies in serum, has higher sensitivity and specificity, greatly improves the detection success rate of ovarian cancer diseases, and has wide application prospects.

Inventors

• LI DAHAI
• WU YINGJUAN

Assignees

• 西安大兴医院

Dates

Publication Date

20260505

Application Date

20260317

Claims (8)

1. 1. The kit for detecting the ovarian cancer is characterized by comprising an ELISA plate and tumor-associated antigens coated on the ELISA plate, wherein the tumor-associated antigens consist of a combination of Sui1, NDE1 and PDLIM 1.
2. 2. The kit for detecting ovarian cancer according to claim 1, wherein the tumor-associated antigen Sui1 coated on the ELISA plate has a coating concentration of 1-2 μg/mL, the tumor-associated antigen NDE1 has a coating concentration of 1-3 μg/mL, and the tumor-associated antigen PDLIM1 has a coating concentration of 3-5 μg/mL.
3. 3. The kit for detecting ovarian cancer according to claim 1, further comprising an enzyme-labeled secondary antibody, a positive control serum, a negative control serum, a washing solution, a color developing solution, and a stop solution.
4. 4. The kit for detecting ovarian cancer according to claim 3, wherein the enzyme-labeled secondary antibody is a horseradish peroxidase-labeled goat anti-human IgG antibody.
5. 5. The kit for detecting ovarian cancer according to claim 3, wherein the positive control serum is a positive control serum of PDLIM1, the negative control serum is a negative control serum of PDLIM1, the washing liquid is a PBST washing liquid, the color developing liquid is a TMB color developing liquid, and the stop solution is sulfuric acid.
6. 6. The kit for detecting ovarian cancer according to claim 5, wherein the positive control serum for PDLIM1 is serum of an ovarian cancer patient positive for PDLIM1 antibody by indirect ELISA and Western blot method, and the negative control serum for PDLIM1 is serum of a healthy person whose PDLIM1 antibody expression level is the average serum antibody content of a healthy person by indirect ELISA and Western blot method.
7. 7. The method according to any one of claims 1 to 6, wherein the method comprises: (1) Preparing an ELISA plate coated with tumor-associated antigens Sui1, NDE1 and PDLIM1, washing the plate by using a washing solution, adding a sample to be detected, incubating for 1-3h at room temperature, and washing by using the washing solution; (2) Adding a goat anti-human IgG antibody marked by horseradish peroxidase into the system in the step (1), incubating for 1-3h at room temperature, and washing with a washing solution; (3) Adding a color development liquid into the system after the reaction in the step (2), developing color for 5-10min at room temperature in a dark place, and adding a stop solution to stop the reaction; (4) OD values at the absorption wavelength of 450 nm were determined using a microplate reader.
8. 8. The method according to claim 7, wherein the sample to be detected is serum.

Description

Kit and detection method for detecting ovarian cancer Technical Field The invention belongs to the technical field of biological medicines, and particularly relates to a kit and a detection method for detecting ovarian cancer. Background Currently, common methods of early screening for ovarian cancer are performed by ovarian cancer tumor markers and pelvic examination. Since about 50% of early ovarian cancer patients have no significant changes in the ovarian cancer tumor marker CA125 found in early detection, it is not ideal in specificity or sensitivity in 90% of patients with advanced disease, whereas pelvic detection is insensitive to detection of ovarian tumor. It is therefore important to find suitable biomarkers to improve the sensitivity and specificity of early screening for ovarian cancer. With the progress of tumor immunotherapy, the relevance of tumors to immunity is becoming more and more important. Research shows that early tumor cells are known to express and produce certain abnormal proteins, which can be recognized by the immune system of the body to produce specific antibodies, wherein the proteins are called tumor-associated antigens, and the antibodies are called autoantibodies. The detection of autoantibodies can indirectly reflect the existence of tumor-associated antigens, and many researches indicate that the autoantibodies generated by tumor antigens are important indexes for early diagnosis of tumors, and can be used for relapse monitoring of tumors and the like. Therefore, the kit for specifically detecting the autoantibody can be used as a complementary means for early detection of the cancer in medicine, and can realize simple, rapid and sensitive detection of the cancer. In general, the diagnosis sensitivity of the individual autoantibodies disclosed at present is low, and the combination of a plurality of autoantibodies can effectively improve the sensitivity, so that new autoantibodies of ovarian cancer are discovered, and a better detection scheme is formed, which has important value for improving the diagnosis accuracy. Based on the above objects, the present invention provides a kit for detecting ovarian cancer and a detection method. Disclosure of Invention The first object of the present invention is to provide a kit for detecting ovarian cancer. In order to achieve the above purpose, the technical scheme adopted by the invention is as follows: the kit for detecting the ovarian cancer comprises an ELISA plate and tumor-associated antigens coated on the ELISA plate, wherein the tumor-associated antigens consist of a combination of Sui1, NDE1 and PDLIM 1. Further, the coating concentration of the tumor-associated antigen Sui1 coated on the ELISA plate is 1-2 mug/mL, the coating concentration of the tumor-associated antigen NDE1 is 1-3 mug/mL, and the coating concentration of the tumor-associated antigen PDLIM1 is 3-5 mug/mL. Further, the kit also comprises enzyme-labeled secondary antibodies, positive control serum, negative control serum, washing liquid, color development liquid and stop solution. Further, the enzyme-labeled secondary antibody is a goat anti-human IgG antibody marked by horseradish peroxidase. Further, the positive control serum is PDLIM1 positive control serum, the negative control serum is PDLIM1 negative control serum, the washing liquid is PBST washing liquid, the color development liquid is TMB color development liquid, and the termination liquid is sulfuric acid. Furthermore, the PDLIM1 positive control serum is the serum of an ovarian cancer patient with positive PDLIM1 antibody detected by using an indirect ELISA and Western blot method, and the PDLIM1 negative control serum is the serum of a healthy person with the average serum antibody content of healthy people with the PDLIM1 antibody expression level detected by using an indirect ELISA and Western blot method. The second object of the present invention is to provide a method for detecting a kit for detecting ovarian cancer. In order to achieve the above purpose, the technical scheme adopted by the invention is as follows: The method for detecting a kit for detecting ovarian cancer according to any one of the above, wherein the method comprises the steps of: (1) Preparing an ELISA plate coated with tumor-associated antigens Sui1, NDE1 and PDLIM1, washing the plate by using a washing solution, adding a sample to be detected, incubating for 1-3h at room temperature, and washing by using the washing solution; (2) Adding a goat anti-human IgG antibody marked by horseradish peroxidase into the system in the step (1), incubating for 1-3h at room temperature, and washing with a washing solution; (3) Adding a color development liquid into the system after the reaction in the step (2), developing color for 5-10min at room temperature in a dark place, and adding a stop solution to stop the reaction; (4) OD values at the absorption wavelength of 450 nm were determined using a microplate reader. Further,