CN111735950A

CN111735950A - Combination of FGF18 and CA125 as early ovarian cancer biomarker and kit

Info

Publication number: CN111735950A
Application number: CN202010694119.9A
Authority: CN
Inventors: 庄光磊; 薛新颖; 刘三宏; 周小进; 师凯旋; 殷霞; 张振峰
Original assignee: Beijing Xinnuo Weikang Technology Co ltd
Current assignee: Beijing Xinnuo Weikang Technology Co ltd
Priority date: 2020-07-17
Filing date: 2020-07-17
Publication date: 2020-10-02
Anticipated expiration: 2040-07-17
Also published as: CN111735950B

Abstract

The invention relates to an early ovarian cancer biomarker and a kit by combining FGF18 and CA125, belonging to the technical field of immunodetection. The invention discloses the use of an agent that detects the levels of FGF18 and CA125 in a sample from a subject in the manufacture of a kit for predicting, assessing or diagnosing early stage ovarian cancer in said subject. The kit provided by the invention can be used for obviously improving the sensitivity and specificity of the prediction, evaluation and diagnosis of ovarian cancer in a subject and greatly reducing the false positive rate in the detection of early ovarian cancer.

Description

Combination of FGF18 and CA125 as early ovarian cancer biomarker and kit

Technical Field

The invention relates to the technical field of immunodetection in general, and particularly relates to a combination of FGF18 and CA125 as an early ovarian cancer biomarker and a kit prepared from the early ovarian cancer biomarker.

Background

Ovarian cancer is one of the common malignant tumors of female reproductive organs, and poses a serious threat to female life. Because the ovary is deeply located in the pelvic cavity, the size is small, and typical symptoms are lacked during the attack, the early diagnosis of the ovarian cancer is difficult, and the technical problem is solved. In clinical findings, it has often progressed to the middle and advanced stages. And progresses to the middle and advanced stages, particularly after metastasis occurs, and the recurrence rate and the 5-year survival rate are low even if the ovarian epithelial cancer is treated by surgery. Therefore, early diagnosis of ovarian cancer is of great significance to reduce metastasis and improve 5-year survival rate. At present, clinically, the diagnosis of ovarian cancer is mainly based on vaginal ultrasonography (TVU) and the detection of ovarian cancer markers in blood

CA-125 (carbohydrate antigen)125, also known as mucin-16, is a protein encoded by the MUC16 gene. CA125 is one of the markers of ovarian epithelial cancer, and is widely used for diagnosis and follow-up of ovarian epithelial cancer at present. However, CA125 is also increased to varying degrees in the case of certain non-malignant diseases, and thus, when CA125 is used to diagnose ovarian cancer, neither specificity nor sensitivity is desirable. In recent years, joint detection using epididymisprotein 4(epididymisprotein 4) and CA125 has been recommended for screening and diagnosis of ovarian cancer (patent documents 1 and 2). However, the sensitivity of single HE4 to the auxiliary diagnosis of stage I ovarian cancer is higher than that of CA125, but is only 45.9%; the sensitivity of The combined detection of HE4 and CA125 was not improved in stage I ovarian cancer (see Moore RG, Brown AK, Miller MC et al, The use of multiple novel tumor biomakers for The detection of ovarian cancer in vitro in antigens with a viral mass. Gynecol Oncol,2008,108: 402-408). Therefore, there is an urgent need to develop new serum markers for early stage ovarian cancer with diagnostic value, so as to determine the occurrence of ovarian cancer by peripheral blood detection sensitively and specifically in the early stage of ovarian cancer.

Human Fibroblast Growth Factor 18(Fibroblast Growth Factor 18, FGF18) is encoded in humans by the FGF18 gene. FGF18 is a member of the Fibroblast Growth Factor (FGF) family. FGF family members have a wide range of mitogenic and cell survival activities and are involved in a variety of biological processes, including embryonic development, cell growth, morphogenesis, tissue repair, tumor growth and invasion. However, the combination of CA125 and FGF18 for biomarkers for early ovarian cancer diagnosis has never been disclosed.

[ REFERENCE ] to

[ patent document 1 ]: CN 108008132A;

[ patent document 2 ]: CN 103954761A.

Disclosure of Invention

The present invention relates generally to early ovarian cancer biomarkers and in particular to methods and uses for predicting, assessing and diagnosing early ovarian cancer by measuring the levels of the biomarker combination FGF18 and CA125, and also provides kits for predicting, assessing and diagnosing early ovarian cancer. The methods and kits provided by the present invention are particularly useful for epithelial ovarian cancer, and more particularly for early stage ovarian cancer (i.e., stage I or II ovarian cancer), thereby providing a method and kit that is easy to handle, highly sensitive, highly specific, and effective for early screening of ovarian cancer. In order to achieve the above purpose of the present invention, the present invention adopts the following technical scheme:

in one aspect, the invention provides the use of an agent that detects the levels of FGF18 and CA125 in a sample from a subject in the manufacture of a kit for predicting, assessing or diagnosing early stage ovarian cancer in said subject. In some embodiments, the ovarian cancer is selected from stage I ovarian cancer, stage II ovarian cancer, or both stage I and stage II ovarian cancer.

In some embodiments, the reagents for detecting FGF18 and CA125 levels are an antibody to FGF18 and an antibody to CA125, respectively. Preferably, the antibody to FGF18 and the antibody to CA125 are a monoclonal antibody to FGF18 and a monoclonal antibody to CA125, respectively.

In some embodiments, the sample is a biological fluid sample from the subject. Preferably, the sample is blood, serum, plasma, lymph, cerebrospinal fluid, ascites, urine and tissue biopsy from the subject. More preferably, the sample is blood, serum and plasma from the subject.

In some embodiments, the kit is a chemiluminescent kit. Preferably, the chemiluminescent kit comprises a capture antibody, a detection antibody and a chemiluminescent substrate.

In some embodiments, the capture antibody is selected from the group consisting of a FGF18 monoclonal antibody linked to a magnetic bead, a CA125 monoclonal antibody linked to a magnetic bead, and a combination thereof. Alternatively, the detection antibody is selected from the group consisting of an FGF18 monoclonal antibody having an alkaline phosphatase label, a CA125 monoclonal antibody having an alkaline phosphatase label, and a combination thereof. Alternatively, the chemiluminescent substrate is AMPPD (CAS No: 122341-56-4).

In another aspect, the invention provides a kit for predicting, assessing or diagnosing ovarian cancer in a subject comprising reagents for detecting FGF18 and CA125 levels in a sample.

Preferably, the ovarian cancer is selected from stage I ovarian cancer, stage II ovarian cancer, or both stage I and stage II ovarian cancer.

In some embodiments, the kit is a chemiluminescent kit. In some embodiments, the kit is a diagnostic kit. Preferably, the chemiluminescent kit comprises a capture antibody, a detection antibody and a chemiluminescent substrate.

In some embodiments, the capture antibody is selected from the group consisting of a FGF18 monoclonal antibody linked to a magnetic bead, a CA125 monoclonal antibody linked to a magnetic bead, and a combination thereof. Alternatively, the detection antibody is selected from the group consisting of an FGF18 monoclonal antibody having an alkaline phosphatase label, a CA125 monoclonal antibody having an alkaline phosphatase label, and a combination thereof. Optionally, the chemiluminescent substrate is AMPPD.

In some embodiments, the kit comprises 2 reagent subgroups, wherein subgroup 1 comprises: magnetic beads coated with monoclonal antibodies against FGF18, FGF18 monoclonal antibodies with alkaline phosphatase label, and the chemiluminescent substrate AMPPD; and is

Wherein subgroup 2 comprises: magnetic beads coated with monoclonal antibodies against CA125, CA125 monoclonal antibodies with alkaline phosphatase label, and the chemiluminescent substrate AMPPD.

In another aspect of the present application, there is also provided a method of predicting, assessing or diagnosing the presence and stage of progression of ovarian cancer in a subject comprising detecting FGF18 and CA125 levels in a sample from the subject. In some embodiments, the level of FGF18 and CA125 in a sample from a subject can be detected by a kit as provided in the above aspects. Alternatively, the detected levels of FGF18 and CA125 in a sample from a subject can be compared to values for FGF18 and CA125 levels obtained from an unaffected population (i.e., standard values) to determine the presence and stage of ovarian cancer in the subject.

Advantageous effects

The present invention uses FGF18 in combination with CA125 as a novel marker for ovarian cancer screening and diagnosis. The present inventors have demonstrated that there is a significant difference in the mean levels of FGF18 and CA125 in serum between the ovarian cancer patient group and the healthy control group (P <0.01), and that there is a correlation between serum FGF18 and CA125 levels in the ovarian cancer patient group (r ═ 0.812). By using the combination of FGF18 and CA125 detection reagents as markers for diagnosing ovarian cancer, compared with the single detection of one of the markers, the sensitivity and specificity of the ovarian cancer diagnosis method and the kit provided by the invention are both obviously improved (the sensitivity is 86.9% & the specificity is 92.1%), the false positive rate is greatly reduced, and a method for screening early ovarian cancer with low misdiagnosis rate, reduced human errors and improved accuracy and a corresponding chemiluminescence detection kit are provided.

Drawings

In order to make the purpose, technical scheme and beneficial effect of the invention more clear, the invention provides the following drawings:

FIG. 1 shows a standard curve for the detection of FGF18 protein by chemiluminescence.

FIG. 2 shows a standard curve for detecting CA125 protein by chemiluminescence method.

Detailed Description

Preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings. Some terms to which the present invention relates are explained as follows.

The biomarker "CA-125", carbohydrate antigen 125, also known as mucin-16, is a protein encoded by the MUC16 gene. CA125 is one of the markers of ovarian epithelial cancer, and is widely used for diagnosis and follow-up of ovarian epithelial cancer at present. Herein, "CA-125" means a polypeptide biomarker or fragment thereof having at least 85% sequence identity to NCBI accession No. NP _ 078966.2.

HUMAN Fibroblast Growth Factor 18(Fibroblast Growth Factor 18, FGF18) is registered in UniProtKB-O76093(FGF18_ HUMAN) in uniprot. FGF18 protein is encoded by the FGF18 gene in humans, is 207 amino acids in length, and is a member of the Fibroblast Growth Factor (FGF) family, which possesses a wide range of mitogenic and cell survival activities, and is involved in a variety of biological processes, including embryonic development, cell growth, morphogenesis, tissue repair, tumor growth, and invasion. The protein was shown to be able to induce neurite outgrowth in PC12 cells in vitro. Studies of similar proteins in mice and chickens have shown that the protein is a pleiotropic growth factor that stimulates proliferation in a variety of tissues, particularly the liver and small intestine. Similar gene knock-out studies in mice suggest that this protein plays a role in regulating the proliferation and differentiation of midline cerebellar structures. Herein, "FGF 18" means a polypeptide biomarker or fragment thereof having at least 85% sequence identity to NCBI accession No. NP _ 003853. Specifically, the full-length amino acid sequence of FGF18 according to accession No. NP _003853 is shown below:

MYSAPSACTCLCLHFLLLCFQVQVLVAEENVDFRIHVENQTRARDDVSRKQLRLYQLYSRTSGKHIQVLGRRISARGEDGDKYAQLLVETDTFGSQVRIKGKETEFYLCMNRKGKLVGKPDGTSKECVFIEKVLENNYTALMSAKYSGWYVGFTKKGRPRKGPKTRENQQDVHFMKRYPKGQPELQKPFKYTTVTKRSRRIRPTHPA。

herein, the types of ovarian cancer predicted, evaluated or diagnosed are serous tumors, endometrioid tumors, mucinous tumors, and clear cell tumors. Also, predicting, assessing or diagnosing ovarian cancer includes predicting specific stages of the disease, such as stage I (IA, IB or IC), stage II, stage III and stage IV tumors. Additionally, "early stage of ovarian cancer" or "early stage ovarian cancer" means ovarian cancer that is in stage I or II. By "diagnosing" is meant identifying the presence or nature of a disorder (i.e., ovarian cancer). Diagnostic methods vary in their sensitivity and specificity. The "sensitivity" of a diagnostic assay is the percentage of diseased individuals in the population that test positive to total diseased individuals. The "specificity" of the diagnostic assay is 1 minus the false positive rate, where the "false positive" rate refers to the proportion of individuals who test positive but are not actually diseased.

"Chemiluminescent kit" means a kit for measuring the level of a biomarker using a Chemiluminescent enzyme immunoassay (cLEIA). Specifically, in the chemiluminescent enzyme immunoassay, an immunoreaction is carried out with an enzyme-labeled bioactive substance, the enzyme on the immunoreactive complex acts on a luminescent substrate, emits light under the action of a signal reagent, and then is subjected to luminescence measurement with a luminescence signal measuring instrument, thereby quantitatively analyzing the level of a biomarker in a sample from a subject.

In some embodiments of the present application, the kits of the present application comprise a capture antibody, a detection antibody, and a chemiluminescent substrate. Specifically, the capture antibody can be prepared by linking a monoclonal antibody against the target biomarker to magnetic beads in the presence of a coupling solution under appropriate reaction conditions. Advantageously, the capture antibody may be obtained by preparing magnetic beads coated with murine anti-human FGF18 monoclonal antibody by coating the carboxylic beads with an appropriate concentration of murine anti-human FGF18 monoclonal antibody in the presence of a coupling solution for 2 hours at 37 ℃ or overnight at 4 ℃.

Further, the detection antibody may be an FGF18 monoclonal antibody with a basic phosphatase label, a CA125 monoclonal antibody with a basic phosphatase label, and combinations thereof. Optionally, the chemiluminescent substrate is AMPPD.

It is noted that although the level of the biomarker in the sample may be detected by other methods known in the art, such as, but not limited to, enzyme linked immunosorbent assay (ELISA), immunofluorescence chromatography, electrochemiluminescence, etc., the present inventors have surprisingly found that when combining chemiluminescence with the biomarkers CA125 and FGF18 of the present application, at least the following advantages are achieved:

1. early stage ovarian cancer patients are detected with high sensitivity (e.g., 10-22 mol/L). This is far above the detection limit of other immunoassay methods, such as radioimmunoassay and enzyme-linked immunoassay.

2. Has wide linear dynamic range. In the methods and kits of the present application, the luminescence intensity is in a linear relationship between 4 and 6 orders of magnitude with the concentration of the substance being measured. This range is much greater than the linear range of absorbance (OD values) in the enzyme immunoassay (2.0).

3. The result is stable, the error is small, the sample directly emits light without any light source irradiation, the influence of a plurality of possible factors (light source stability, light scattering, optical wave selector and the like) on the analysis is eliminated, and the analysis is sensitive, stable and reliable.

Thus, the combination of chemiluminescence with the biomarkers CA125 and FGF18 of the present application has the advantages described above.

Examples

The purchase information of the products used in the examples is as follows, but is not limited to the same manufacturer.

Mouse anti-human FGF18 monoclonal antibody (coating antibody): the goods number is: sc-393471, available from Santacruz under the trade name FGF-18 antibody.

Mouse anti-human FGF18 monoclonal antibody (detection antibody): the goods number is: ab86571 was purchased from Abcam under the trade name Anti-FGF18 antibody.

FGF18 protein standard: the goods number is: 13206-H08H, available from SinoBiological under the trade name FGF18 protein.

Mouse anti-human CA125 monoclonal antibody (coating antibody), cat #: CA125-McAb2, purchased from Fipeng organisms under the trade name carbohydrate antigen 125 antibody.

Mouse anti-human CA125 monoclonal antibody (detection antibody), cat #: CA125-McAb1, purchased from Fipeng organisms under the trade name carbohydrate antigen 125 antibody.

Standard CA125 protein: the goods number is: CA125-Ag, purchased from Fipeng organisms under the trade name carbohydrate antigen 125.

Magnetic beads: the goods number is: MagCOOH, commercially available from knoyi microsphere technologies, inc.

Alkaline phosphatase (alkaline phosphatase): the goods number is: SP011401, available from national reagent under the trade name alkaline phosphatase.

AMPPD: CAS No. 122341-56-4, 4-methoxy-4- (3-phosphorylphenyl) spiro [1, 2-dioxetane-3, 2' -adamantane ], is an alkaline phosphatase substrate, commercially available from kyoto jenko jiekang under the trade name chemiluminescence substrate.

Tween-20: tween-20 is available from biologies under the trade name.

Example 1 establishment of FGF18 detection System and optimization thereof

Constructing a detection system: coating the carboxyl magnetic beads with the mouse anti-human FGF18 monoclonal antibody with the concentration of 2.5 mug/mL at 37 ℃ for 2 hours or at 4 ℃ overnight to prepare magnetic beads coated with the mouse anti-human FGF18 monoclonal antibody, namely capture antibodies; then adding FGF18 protein standard and serum samples with the concentrations of 1000, 500, 250, 125, 62.5, 31.25, 15.625 and 0ng/ml into different reaction cups respectively, and reacting for 30 minutes at 37 ℃; then adding a mouse anti-human FGF18 monoclonal antibody (namely, a detection antibody) marked by alkaline phosphatase with the concentration of 0.25 mu g/mL, reacting for 30 minutes at 37 ℃, washing magnetic beads, and removing supernatant; finally, a luminescent substrate, AMPPD, was added and the luminescence value was measured. An FGF18 protein standard curve was constructed based on the luminescence values of the reaction system obtained from the standard values of different concentrations, and the results are shown in FIG. 1. As can be seen from FIG. 1, the linear range of the FGF18 detection system is 0ng/mL-1000ng/mL, the linear correlation coefficient r of the standard substance in the linear range is more than or equal to 0.990, and the recovery rate is in the range of 90% -110%.

Determining a detection system: the antibody with different concentrations is detected by a checkerboard matrix method, wherein the capture antibody is selected from 0.5, 1.5, 2.5, 3.5, 4.5 and 5.5 mu g/mL, and the detection antibody is selected from 0.05, 0.15, 0.2, 0.25, 0.35, 0.45 and 0.55 mu g/mL. The antibody with different concentrations is detected, and the optimal working concentration of the FGF18 capture antibody is found to be 2.5 mu g/mL, and the optimal working concentration of the FGF18 detection antibody is found to be 0.25 mu g/mL.

Example 2 FGF18 chemiluminescent assay kit

An FGF18 chemiluminescence assay kit is constructed according to an FGF18 serum assay system established in example 1, and the specific components are shown in Table 1:

TABLE 1 Components of FGF18 chemiluminescent assay kit

Evaluation of FGF18 chemiluminescent detection kit: detecting the FGF18 positive quality control product by using an FGF18 chemiluminescence detection kit, and repeatedly detecting for 10 times at two levels of FGF18 protein concentration of 62.5ng/mL and FGF18 protein concentration of 500ng/mL respectively, wherein the result shows that the coefficient of variation CV is less than or equal to 10%; when 3 batch number kits are used for detecting the same sample, the inter-batch variation coefficient CV of the 3 batch number kits is less than or equal to 12 percent.

Example 3 establishment of CA125 serum detection reaction System and optimization thereof

Coating the magnetic beads with a mouse anti-human CA125 monoclonal antibody with the concentration of 5 mu g/mL, and coating overnight at 37 ℃ for 2 hours or 4 ℃; respectively adding the CA125 protein standard substance and the serum sample with the concentrations of 0U/mL, 10U/mL, 20U/mL, 40U/mL, 80U/mL, 160U/mL and 320U/mL into the reaction holes, and reacting for 30 minutes at 37 ℃; then, a mouse anti-human CA125 monoclonal antibody marked by alkaline phosphatase with the concentration of 0.5 mu g/mL is used for reacting for 30 minutes at the temperature of 37 ℃, and the magnetic beads are washed to remove the supernatant; finally, a luminescent substrate, AMPPD, was added and the luminescence value was measured. A calibration curve of CA125 was constructed based on the luminescence values of the reaction system obtained from the standard values of different concentrations, and the results are shown in FIG. 2. As can be seen from FIG. 2, the linear range of the CA125 chemiluminescence detection kit is 10U/mL-320U/mL, the linear correlation coefficient r of the standard substance in the linear range is more than or equal to 0.990, and the recovery rate is in the range of 90% -110%.

Determining a detection system: similar to the checkerboard method in example 2, the best working concentration of the home-made CA125 capture antibody was found to be 5. mu.g/mL and the best working concentration of the CA125 detection antibody was found to be 0.5. mu.g/mL by detecting different concentrations of the antibody.

The main components of the detection system are determined through the research, and then a CA125 serum detection system is established.

Example 4 CA125 chemiluminescence detection kit

The CA125 chemiluminescence assay kit is constructed according to the CA125 serum assay system established in the example 3, and the specific components are shown in the following table 2:

TABLE 2 CA125 chemiluminescence assay kit Components

Evaluation of CA125 chemiluminescent assay kit: detecting a CA125 repetitive reference substance by using a CA125 chemiluminescence detection kit, and repeatedly detecting for 10 times at two levels of 20U/mL and 80pg/mL of CA125 protein concentration respectively, wherein the result shows that the coefficient of variation CV is less than or equal to 10%; when 3 batch number kits are used for detecting the same sample, the inter-batch variation coefficient CV of the 3 batch number kits is less than or equal to 15 percent.

Example 5 Combined detection kit for human ovarian cancer marker FGF18-CA125

The FGF18 chemiluminescence detection kit constructed in example 2 and the CA125 chemiluminescence detection kit constructed in example 4 are combined to form an FGF18-CA125 combined detection kit.

Example 6 FGF18-CA125 Combined detection kit diagnosis and prognosis of early ovarian cancer

100 clinical samples were collected, 22 samples from the normal population and 78 samples from early stage ovarian cancer patients, each with 1mL serum. Of the 78 ovarian cancer patients, 24 were stage I ovarian cancer patients and 54 were stage II ovarian cancer patients. Respectively detecting the concentrations of FGF18 and CA125 markers in blood serum of ovarian cancer patients and healthy normal people, analyzing the obtained data by using SPASS statistical software according to the detection result, and if the difference between different groups has statistical significance, performing independent sample t test, wherein the result is expressed by x +/-s; correlation analysis was performed between FGF18 and CA125 in each group, with P <0.05 as the difference being statistically significant. Finally, the specificity and sensitivity of the FGF18 and CA125 markers were determined individually or in combination according to data statistics, and the results are shown in Table 3, with serum FGF18 (251. + -. 114.51) ng/mL in ovarian cancer group; CA125(591.96 + -384.5) U/mL, both higher than healthy control FGF18(132 + -83.82) ng/mL; CA125 (30.45. + -. 25.87) U/mL, statistically treated, and the difference was statistically significant (p < 0.01).

TABLE 3 serum FGF18 and CA125 assay results (X + -S) for ovarian cancer group and healthy control group

Group of	n	Serum FGF18(ng/mL)	CA125(U/mL)
				Healthy control group	22	132±83.82	30.45±25.87
Ovarian cancer	78	251±114.51	591.96±384.5

Note: compared with the healthy control group, the ovarian cancer group has P less than 0.05; comparison of serum FGF18 between ovarian cancer and healthy controls, P < 0.05; comparison of serum CA125 levels between ovarian cancer and healthy controls, P < 0.05.

Meanwhile, there was a correlation (r ═ 0.812) between the levels of FGF18 and CA125 in the ovarian cancer group, whereas there was no significant correlation (r ═ 0.017) between the levels of FGF18 and CA125 in the healthy control group, and the results are shown in table 4.

TABLE 4 correlation between FGF18 and CA125 in ovarian cancer and healthy control groups

Group of	n	Serum FGF18(ng/mL)	CA125(U/mL)	Coefficient of correlation r
					Healthy control group	22	132±83.82	30.45±25.87	0.017
Ovarian cancer	78	251±114.51	591.96±384.5	0.812

Note: there was a significant correlation between the levels of CA125 and FGF18 in the ovarian cancer group (r 0.812), while there was no significant correlation between the levels of CA125 and FGF18 in the healthy control group (r 0.017).

Further, by analyzing the sensitivity and specificity of the ovarian cancer diagnosis cases, the following results can be obtained: the sensitivity of the FGF18 marker detected alone was 72.3%, and the specificity was 85.4%; the sensitivity of the CA125 marker alone was 82.5% and the specificity was 70.1%. The sensitivity of the combined assay of the two markers FGF18 and CA125 was 86.9% and the specificity was 92.1% (see table 5 below). Therefore, the combined detection of the two markers of FGF18 and CA125 is obviously superior to the detection of a single ovarian cancer marker.

TABLE 5 ovarian cancer diagnosis results of FGF18-CA125 combined detection kit

Marker substance	Area under curve	Sensitivity of the probe	Specificity of
				FGF18	0.792	72.3％	85.4％
CA125	0.834	82.5％	70.1％
				FGF18+CA125	0.892	86.9％	92.1％

Note: the combined detection of the two markers of FGF18 and CA125 is obviously superior to the detection of a single ovarian cancer marker in terms of sensitivity and specificity.

In conclusion, FGF18 and CA125 can be used as markers for diagnosing and predicting early ovarian cancer, can be used for early diagnosis of ovarian cancer, and improves accuracy of early prediction.

Finally, it is noted that the above-mentioned preferred embodiments illustrate rather than limit the invention, and that, although the invention has been described in detail with reference to the above-mentioned preferred embodiments, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the scope of the invention as defined by the appended claims.

Claims

1. Use of an agent that detects the levels of FGF18 and CA125 in a sample from a subject, wherein the ovarian cancer is selected from stage I ovarian cancer, stage II ovarian cancer, or both stage I and stage II ovarian cancer, in the manufacture of a kit for predicting, assessing or diagnosing ovarian cancer in the subject, wherein the sample is blood, serum, plasma from the subject.

2. The use according to claim 1, wherein the reagents for detecting the levels of FGF18 and CA125 are an antibody directed against FGF18 and an antibody directed against CA125, respectively.

3. The use according to claim 2, wherein the antibody directed to FGF18 and the antibody directed to CA125 are a monoclonal antibody directed to FGF18 and a monoclonal antibody directed to CA125, respectively.

4. Use according to claim 1, wherein the kit is a chemiluminescent kit comprising a capture antibody, a detection antibody and a chemiluminescent substrate.

5. The use of claim 4, wherein the capture antibody is selected from the group consisting of an FGF18 monoclonal antibody linked to a magnetic bead, a CA125 monoclonal antibody linked to a magnetic bead, and combinations thereof;

the detection antibody is selected from the group consisting of an FGF18 monoclonal antibody with an alkaline phosphatase label, a CA125 monoclonal antibody with an alkaline phosphatase label, and combinations thereof; and/or the presence of a gas in the gas,

the chemiluminescent substrate is AMPPD.

6. A kit for predicting, assessing or diagnosing ovarian cancer in a subject, comprising reagents for detecting FGF18 and CA125 levels in a sample,

wherein the ovarian cancer is selected from stage I ovarian cancer, stage II ovarian cancer, or both stage I and stage II ovarian cancer, and the sample is blood, serum, plasma from the subject.

7. The kit of claim 6, wherein the reagents for detecting the levels of FGF18 and CA125 are an antibody directed against FGF18 and an antibody directed against CA125, respectively.

8. The kit of claim 7, wherein the antibody to FGF18 and the antibody to CA125 are a monoclonal antibody to FGF18 and a monoclonal antibody to CA125, respectively.

9. The kit of claim 6, wherein the kit is a chemiluminescent kit comprising a capture antibody, a detection antibody, and a chemiluminescent substrate, wherein: the capture antibody is selected from the group consisting of an FGF18 monoclonal antibody linked to a magnetic bead, a CA125 monoclonal antibody linked to a magnetic bead, and combinations thereof;

the chemiluminescent substrate is AMPPD.

10. The kit of claim 6, wherein the kit comprises 2 reagent subgroups,

wherein subgroup 1 comprises: magnetic beads coated with monoclonal antibodies against FGF18, FGF18 monoclonal antibodies with alkaline phosphatase label, and the chemiluminescent substrate AMPPD; and is