Kit for screening cervix canceration and application thereof

Abstract

The invention belongs to the technical field of biomedicine, particularly relates to the field of molecular diagnosis, and more particularly relates to a kit for screening cervical canceration and application of the kit. The invention provides a biomarker for screening cervix canceration, the biomarker is at least one of an XIRP2 autoantibody, a POU3F2 autoantibody and a PCNA autoantibody, the expression level of the marker in serum of a patient with cervix canceration is obviously higher than that of a normal person, and the difference has statistical significance. The invention also provides a kit for screening cervical canceration. The kit contains an antigen for detecting the biomarker. By detecting the expression level of the biomarker in human serum, the biomarker can be used for screening and diagnosing cervical canceration.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biomedical technology, in particular to the field of molecular diagnosis, and more particularly to a kit for screening cervical cancer and application thereof. BACKGROUND

[0002] Cervical cancer is the fourth most common cancer in women worldwide, and its development usually goes through a continuous process from low-grade squamous intraepithelial lesion (LSIL) to high-grade squamous intraepithelial lesion (HSIL II / III) and then to invasive cancer.

[0003] The key to eliminating cervical cancer is high-quality screening. The main methods for cervical cancer screening currently include cervical cytology examination (such as Pap smear and liquid-based cytology test (TCT)) and human papillomavirus (HPV) detection. However, cytology examination has strong subjectivity and insufficient sensitivity, and its diagnostic efficiency still needs to be improved. Although HPV DNA detection has high sensitivity, its specificity is low because most HPV infections are transient and do not progress to precancerous lesions or cancer, which easily leads to a large number of unnecessary colposcopy referrals and patient anxiety. Therefore, it is urgent to develop molecular markers with higher specificity for accurately identifying high-risk lesions that have the risk of progression. SUMMARY

[0004] In view of the problems such as insufficient specificity in the existing cervical cancer screening technologies, the present application provides a kit for screening cervical cancer and application thereof.

[0005] To achieve the above purpose, the present application adopts the following technical solutions:

[0006] The present application provides, in a first aspect, an application of a reagent for detecting a biomarker in the preparation of a product for screening cervical cancer, wherein the biomarker is at least one of XIRP2 autoantibody, POU3F2 autoantibody and PCNA autoantibody.

[0007] According to the above application, preferably, the cervical cancer is cervical intraepithelial neoplasia (CIN) HSIL II / III or cervical cancer.

[0008] According to the above application, preferably, the reagent is a reagent for detecting the biomarker in a sample by enzyme-linked immunosorbent assay, protein chip, immunoblotting or microfluidic immunoassay.

[0009] According to the above application, preferably, the reagent is an antigen or an antibody for detecting the biomarker. More preferably, the reagent is an antigen for detecting the biomarker, and the antigen is at least one of XIRP2 protein, POU3F2 protein and PCNA protein.

[0010] According to the above-mentioned application, preferably, the sample is serum, plasma, interstitial fluid or cervical exfoliated cell sample.

[0011] According to the above-mentioned application, preferably, the product is a protein chip, a kit or a preparation.

[0012] The second aspect of the present application provides a kit for screening cervical canceration, preferably, the kit comprises reagents for detecting biomarkers, the biomarkers being at least one of XIRP2 autoantibody, POU3F2 autoantibody and PCNA autoantibody.

[0013] According to the above-mentioned kit, preferably, the kit detects the biomarkers in the sample by enzyme-linked immunosorbent assay, protein chip, immunoblotting or microfluidic immunoassay.

[0014] According to the above-mentioned kit, preferably, the kit is an ELISA detection kit, which comprises a solid carrier and an antigen coated on the solid carrier; the antigen is at least one of XIRP2 protein, POU3F2 protein and PCNA protein.

[0015] According to the above-mentioned kit, preferably, the sample detected by the kit is serum, plasma, interstitial fluid or cervical exfoliated cell sample.

[0016] According to the above-mentioned kit, preferably, the ELISA detection kit further comprises sample diluent, secondary antibody, antibody diluent, washing solution, color developing solution and termination solution.

[0017] The basic information of XIRP2 protein, POU3F2 protein and PCNA protein in the present application is as follows, for example:

[0018] XIRP2 (NP_689594.4) is a member of the actin-binding, actin repeat-containing protein family. Under normal physiological conditions, XIRP2 protein is highly expressed in skeletal muscle and cardiac muscle cells, and plays a protective role in preventing actin filament depolymerization. Studies have found that XIRP2 mutation is a common mutation (> 3%) in cancer and is associated with negative prognosis. Currently, high frequency mutations of XIRP2 have been reported to be associated with the occurrence and prognosis of high-grade neuroblastoma, breast cancer and primary liver cancer.

[0019] POU3F2 (NP_005595.2) is a transcription factor that plays a key role in neural development. In recent years, studies have found that POU3F2 is abnormally expressed in various malignant tumors, and is closely related to poor prognosis and treatment resistance as an important cancer gene. Studies have shown that high expression of POU3F2 is an independent influencing factor for poor prognosis of patients with melanoma, breast cancer and endometrial cancer.

[0020] PCNA (NP_872590.1) is a key marker of cell proliferation, closely related to DNA damage repair and cell cycle regulation, and its activated form is enriched in cells with active proliferation, such as cancer cells. The function and expression of PCNA are regulated at multiple levels, including post-transcriptional and post-translational modifications. For example, the up-regulation of certain long non-coding RNAs (lncRNAs), such as in esophageal squamous cell carcinoma, can promote tumor progression by regulating PCNA.

[0021] Compared with the prior art, the present application has the following positive and beneficial effects:

[0022] (1) The present application first found that the expression levels of XIRP2 autoantibody, POU3F2 autoantibody and PCNA autoantibody in the serum of cervical cancer patients were significantly higher than those in the healthy control group. Therefore, the present application proposes to develop a cervical cancer diagnostic kit based on these three autoantibodies. When using XIRP2 autoantibody to diagnose and distinguish cervical cancer patients from normal people, the AUC is 0.76; when using POU3F2 autoantibody to diagnose and distinguish cervical cancer patients from normal people, the AUC is 0.74; when using PCNA autoantibody to diagnose and distinguish cervical cancer patients from normal people, the AUC is 0.69. Compared with a single specific antigen autoantibody, the AUC and sensitivity of multiple specific antigen autoantibodies used for cervical cancer diagnosis are significantly improved. When PCNA+POU3F2 is used to distinguish cervical cancer patients from normal people, the AUC is 0.81; when XIRP2+PCNA is used to distinguish cervical cancer patients from normal people, the AUC is 0.84; when XIRP2+POU3F2 is used to distinguish cervical cancer patients from normal people, the AUC is 0.85; when XIRP2+POU3F2+PCNA is used to distinguish cervical cancer patients from normal people, the AUC reaches a maximum of 0.92, the sensitivity of detection reaches 42.9%, and has a high diagnostic value, while the specificity of detection can still reach 93.0%.

[0023] (2) The kit prepared by the present application can accurately distinguish cervical cancer patients from healthy controls by detecting the expression levels of XIRP2 autoantibody, POU3F2 autoantibody and PCNA autoantibody in human serum by indirect ELISA method, which provides a new reference for the diagnosis of cervical cancer for clinicians. The kit is expected to improve the early diagnosis level of cervical cancer screening in China, and provide a new direction for the mechanism research and diagnosis and treatment of cervical cancer. BRIEF DESCRIPTION OF DRAWINGS

[0024] Figure 1 is the expression level of the autoantibody of the three specific antigens (XIRP2, POU3F2, PCNA) in the serum of the cervical cancer group.

[0025] Figure 2 The result chart of the expression level of autoantibodies for three specific antigens (XIRP2, POU3F2, PCNA) in the control group serum;

[0026] Figure 3 The result chart of the positive rate of autoantibodies for three specific antigens (XIRP2, POU3F2, PCNA) in the cervical cancer group and the control group;

[0027] Figure 4 The ROC curve of autoantibodies for three specific antigens (XIRP2, POU3F2, PCNA) in the single diagnosis of distinguishing cervical cancer patients and normal people;

[0028] Figure 5 The ROC curve of autoantibodies for three specific antigens (XIRP2, POU3F2, PCNA) in the combined diagnosis of cervical cancer. DETAILED DESCRIPTION

[0029] In order to make the objects, technical solutions and advantages of the present application clearer, the present application will be further described in detail through examples. It should be understood that the specific examples described herein are only used to explain the present application, and are not used to limit the present application.

[0030] The present inventors screened specific antigen autoantibodies differentially expressed in the serum of cervical cancer patients (including HSIL II / III grade and cervical cancer) and healthy controls by using HuProtTM human proteome chip in the early stage, which are XIRP2 autoantibody, POU3F2 autoantibody and PCNA autoantibody. The expression levels of XIRP2 autoantibody, POU3F2 autoantibody and PCNA autoantibody in the serum of cervical cancer patients are significantly higher than those in the serum of healthy controls, and the difference has statistical significance.

[0031] In order to verify the difference of the expression levels of the above three autoantibodies, the expression levels of the screened XIRP2 autoantibody, POU3F2 autoantibody and PCNA autoantibody were further detected in the serum of a large sample population by indirect enzyme-linked immunosorbent assay using ELISA kit.

[0032] Example 1: Preparation of ELISA detection kit

[0033] 1. Experimental materials and reagents:

[0034] (1) Three specific antigen proteins: XIRP2 protein, POU3F2 protein and PCNA protein were purchased from Genesci (Shanghai) Co., Ltd. or Beijing Zhongshanjingqiao Biotechnology Co., Ltd. and other suppliers;

[0035] (2) 96-well enzyme-labeled plate (8 rows x 12 columns);

[0036] (3) Coating solution: 50 mM carbonate buffer, pH = 9.6;

[0037] (4) Blocking solution: PBST buffer containing 2% (v / v) bovine serum albumin (BSA) and 0.2% (v / v) Tween 20;

[0038] (5) Sample diluent: PBST buffer containing 1% (W / V) BSA;

[0039] (6) Secondary antibody diluent: PBST buffer containing 1% (W / V) BSA;

[0040] (7) Enzyme-labeled secondary antibody: horseradish peroxidase-labeled RecA protein (e.g. Invitrogen Corporation);

[0041] (8) Washing solution: PBST (phosphate buffered Tween) buffer containing 0.05% Tween 20, pH 7.4;

[0042] (9) Positive control serum: XIRP2 positive control serum, i.e. serum from a cervical cancer patient that is positive for XIRP2 antibodies as detected by indirect ELISA and Western blot methods;

[0043] (10) Negative control serum: XIRP2 negative control serum, i.e. serum from a normal person whose XIRP2 antibody expression level is equal to the average content of serum antibodies in the normal population;

[0044] (11) Color developing solution A: 0.02% (W / V) TMB, prepared by dissolving 0.005 g of methylbenzidine (TMB) in 25 mL of deionized water;

[0045] (12) Color developing solution B: 0.006% (W / V) urea peroxide, prepared by dissolving 4.665 g of citric acid and 18.40 g of Na2HPO4 in 400 mL of deionized water, adding 0.75% urea peroxide 3.2 mL, adjusting the pH to 5.0-5.5, and diluting with deionized water to a final volume of 500 mL, and mixing well at 4°C;

[0046] (13) Stop solution: 10% sulfuric acid;

[0047] (14) Enzyme-labeled plate reader: Star Fax 2100 (Awareness, USA).

[0048] 2. Preparation of antigen-coated enzyme-labeled plate:

[0049] Prepare the enzyme-labeled plate coated with specific antigen XIRP2, the enzyme-labeled plate coated with POU3F2, and the enzyme-labeled plate coated with PCNA, respectively.

[0050] Take the preparation of the enzyme-labeled plate coated with antigen XIRP2 as an example, and the specific operation steps are as follows:

[0051] (1) Prepare a specific antigen solution: prepare a XIRP2 protein solution with a concentration of 0.125 g / mL by using a coating solution to coat the specific antigen XIRP2 protein;

[0052] (2) Coat the enzyme-labeled plate: add the specific antigen solution (XIRP2 protein solution) prepared in step 1) to each reaction well of the 96-well enzyme-labeled plate, with a sample amount of 50 L / well; add XIRP2 antigen solution (with a sample amount of 50 L / well) to the positive control well and the negative control well, and add the coating solution (with a sample amount of 50 L / well) to the blank control well; incubate in a 37℃ constant temperature incubator for 1 h, remove the coating solution after overnight incubation at 4℃, and then wash with the washing solution for 3 times, each time for 3 min;

[0053] (3) Blocking: add blocking solution to the reaction wells of the coated 96-well enzyme-labeled plate, with a sample amount of 100 L / well, and block in a 37℃ water bath for 2 h; then remove the blocking solution, wash with the washing solution (with a sample amount of 300 L / well) for 3 times, and pat dry to obtain the enzyme-labeled plate coated with specific related antigen XIRP2;

[0054] The operation steps for preparing the enzyme-labeled plate coated with antigen POU3F2 and the enzyme-labeled plate coated with antigen PCNA are basically the same as those for preparing the enzyme-labeled plate coated with XIRP2, and the coating concentrations are also the same. The difference lies in that the specific related antigens used in step 1) are different, and the specific antigen solutions added to the reaction wells of the 96-well enzyme-labeled plate in step 2) are different. When preparing the enzyme-labeled plate coated with antigen POU3F2, the specific antigen used in step 1) is POU3F2 protein, and the specific antigen solution added to the reaction wells of the 96-well enzyme-labeled plate in step 2) is POU3F2 protein solution. When preparing the enzyme-labeled plate coated with antigen PCNA, the specific antigen used in step 1) is PCNA protein, and the specific antigen solution added to the reaction wells of the 96-well enzyme-labeled plate in step 2) is PCNA protein solution.

[0055] 3, The composition of the kit of the application is as follows:

[0056] (1) The 96-well enzyme-labeled plate coated with antigens (XIRP2, POU3F2, PCNA, or combined as needed) prepared in step 3;

[0057] (2) Sample diluent: PBST buffer containing 1% (W / V) BSA;

[0058] (3) Second antibody diluent: PBST buffer containing 1% (W / V) BSA;

[0059] (4) Enzyme-labeled second antibody: horseradish peroxidase-labeled RecA protein;

[0060] (5) Color developing solution: the color developing solution is composed of color developing solution A and color developing solution B, wherein the color developing solution A is 0.02% (W / V) TMB, and the color developing solution B is 0.006% (W / V) urea peroxide; when used, the color developing solution A and the color developing solution B are mixed in equal volumes at a ratio of 1:1.

[0061] (6) Stop solution: 10% sulfuric acid;

[0062] (7) Washing solution: 0.01M PBST (phosphate buffered saline Tween) buffer containing 0.05% Tween 20 at pH 7.4;

[0063] (8) Positive control serum: XIRP2 positive control serum (or selected according to the marker);

[0064] (9) Negative control serum: XIRP2 negative control serum (or selected according to the marker).

[0065] The reagents (2) to (9) are packaged respectively to constitute a kit with the 96-well enzyme-labeled plate coated with the antigen.

[0066] Example 2: Use of the ELISA detection kit

[0067] 1. Incubation of serum samples:

[0068] The serum sample to be detected is diluted by 1:100 in volume ratio with the serum sample diluent. The diluted serum sample is added to the reaction wells of columns 1 to 11 of the 96-well enzyme-labeled plate coated with the XIRP2 protein prepared in the above step (1) at an amount of 50 μL / well; the positive serum diluted at a volume ratio of 1:100 is added to the first to third reaction wells of column 12 of the 96-well enzyme-labeled plate coated with the XIRP2 protein at an amount of 50 μL / well, and the negative serum diluted at a volume ratio of 1:100 is added to the fourth to sixth reaction wells of column 12 at an amount of 50 μL / well; the antibody diluent without serum (at an amount of 50 μL / well) is added to the seventh to eighth reaction wells of column 12 of the 96-well enzyme-labeled plate coated with the XIRP2 protein as a blank control; then the 96-well enzyme-labeled plate is incubated in a 37°C water bath for 1 h, and then the liquid in the reaction wells is discarded, washed with the washing solution (at an amount of 300 μL / well) for 5 times, and patted dry.

[0069] 2. Incubation of secondary antibody:

[0070] The horseradish peroxidase-labeled RecA protein is diluted with the second antibody diluent at a ratio of 1:40000, and then the diluted horseradish peroxidase-labeled RecA protein is added to the reaction wells of the 96-well enzyme-labeled plate at a sample amount of 50 μL / well, and incubated in a 37°C water bath for 1 h, and then the liquid in the reaction wells is discarded, washed with a washing solution (a sample amount of 300 μL / well) for 5 times and patted dry.

[0071] 3. Color development and termination reaction:

[0072] The color developing solution A and the color developing solution B are mixed uniformly at a ratio of 1:1 by volume, and then the mixed color developing solution is rapidly added to the reaction wells of the 96-well enzyme-labeled plate at a sample amount of 50 μL / well, and color development reaction is carried out at room temperature in the dark for 5-15 min, and then 25 μl of a termination solution is further added to each reaction well to terminate the color development reaction; and an enzyme-labeled instrument is used to read the absorbance OD450 and OD620 at 450 nm and 620 nm, respectively, wherein the absorbance OD620 at 620 nm is a background value, and the difference between OD450 and OD620 is taken as the final result of the detected absorbance value.

[0073] The specific operation steps for detecting the expression levels of POU3F2 autoantibodies and PCNA autoantibodies in serum samples are basically the same as those for detecting XIRP2 autoantibodies, and the difference lies in that in step 1), the enzyme-labeled plates used for detection are POU3F2 and PCNA protein-coated enzyme-labeled plates, respectively.

[0074] 4. Data processing:

[0075] The absorbance values (OD values) of the serum samples of the cervical cancer group and the normal control group are subjected to Kolmogorov-Smirnova test, and the results show that the expression levels of the three specific related antigen autoantibodies in the serum samples of the research objects do not conform to the normal distribution (P<0.05), and therefore the 25th percentile (P25), the median (P50) and the 75th percentile (P75) are used to describe the expression level distribution of the three specific antigen autoantibodies; and then a non-parametric test (Mann-Whitney U) is used to compare whether there is a difference in the expression levels of the autoantibodies in the cervical cancer group and the normal control group.

[0076] 5. Determination of the detection results of the kit:

[0077] The average of the OD values measured by the negative control holes plus two standard deviations (Mean+2SD) is taken as the cut-off value, and the OD value reading in the reaction well is judged as positive if it is greater than or equal to the cut-off value, and is judged as negative if it is less than the cut-off value.

[0078] Example 3 Analysis of the diagnostic value of the kit of the present application

[0079] 1. Experimental samples:

[0080] Serum samples from 112 patients with cervical cancer (HSIL (CIN II / III) and cervical cancer) and 100 healthy people (control group) were selected, including 80 cases of HSIL (denoted as the HSIL group), with an average age of 45.0 ± 1.4 years (20-76 years old); among the 112 patients in the cervical cancer group, there were 32 cases of cervical cancer (denoted as the cervical cancer group), with an average age of 51.3 ± 9.5 years (32-70 years old). The average age of the 100 cases in the control group was 58.8 ± 1.2 years (30-79 years old), and they were all from the hospital's healthy examination population, with normal cervical screening results (cytology and HPV detection were both negative) and no history of cervical lesions. This study was approved by the hospital ethics committee, and all the research subjects had signed the informed consent form.

[0081] Serum collection: 5 ml of peripheral blood of the research subjects in the morning on an empty stomach was collected in a blood collection tube without anticoagulant, and then placed at room temperature for 1 h, and then placed in a centrifuge, set at 4°C, 3000 rpm for 10 min. Then the serum on the top of the blood collection tube was sucked out and divided into 1.5 mL EP tubes, the sample number was marked on the top and side of the EP tube, and then placed in a-80°C refrigerator for frozen storage, and the blood collection date and storage location were recorded. Before use, the serum was taken out and placed in a 4°C refrigerator for thawing and sub-packaging to avoid repeated freezing and thawing of the serum.

[0082] 2. Experimental method:

[0083] The expression levels of the three specific antigen autoantibodies in the serum samples of the cervical cancer group, the HSIL group and the control group were detected by using the ELISA kit prepared in Example 1 and the use method of the ELISA kit described in Example 2. The same serum sample was used to detect the expression levels of XIRP2 autoantibody, POU3F2 autoantibody and PCNA autoantibody in the serum sample by using the above-prepared ELISA kit.

[0084] 3. Experimental results:

[0085] (1) The expression levels of the three specific antigen autoantibodies in the cervical cancer group, the HSIL group and the control group:

[0086] The statistical results of the expression levels of the three specific antigen autoantibodies in the cervical cancer group, the HSIL group and the control group are shown in Figure 1 , Figure 2 and Figure 3 , and the expression level OD value is shown in Table 1. From Figure 1 and Figure 3It can be seen that the average expression level of the three specific antigen autoantibodies in the cervical cancer group is significantly higher than that in the control group. Figure 1 、 Figure 2 and Figure 3 It can be seen that the average expression level of the three specific antigen autoantibodies in the control group, HSIL group and cervical cancer group increases (see Table 1), indicating that the three specific antigen autoantibodies can be used for the diagnosis of cervical cancer.

[0087] Table 1 is the average expression level (OD value, M ± SD) of the three specific antigen autoantibodies in the cervical cancer group, HSIL group and control group

[0088]

[0089] (2) The positive rate of the three specific antigen autoantibodies in the cervical cancer group, HSIL group and control group:

[0090] The positive rate of the three specific antigen autoantibodies in the cervical cancer group, HSIL group and control group was calculated according to the detection results of the above kit (the number of positive objects detected in each group divided by the total number of objects detected in the group), and the bar chart of the positive rate of the three tumor-related antigen autoantibodies in the cervical cancer group and the normal control group was drawn using Excel software, as shown in Figure 4 The SPSS 29.0 software was used for statistical test, and the two independent sample chi-square test method was used to compare the antibody positive rates of the cervical cancer group, HSIL group and control group, with a test level α = 0.05. When P < 0.05, the result was statistically significant. It can be seen from Figure 4 that the positive rate of the three specific antigen autoantibodies in the serum of the cervical cancer group is significantly higher than that in the HSIL group, and the HSIL group is higher than the control group. Therefore, the above three specific antigen autoantibodies can be used as detection indicators for cervical cancer, and can be used for early diagnosis of cervical cancer.

[0091] (3) Evaluation of the value of the three specific antigen autoantibodies in the diagnosis of cervical cancer:

[0092] Further, according to the levels of the three specific antigen autoantibodies in the cervical cancer group, HSIL group and control group detected by the kit, the ROC curve of the three tumor-related antigen autoantibodies for diagnosing and distinguishing cervical cancer patients and normal people was drawn using GraphPad Prism 8.0, the value of the three specific antigen autoantibodies for diagnosing and distinguishing cervical cancer patients and normal people was verified, and the corresponding sensitivity and specificity were calculated.

[0093] Because the sensitivity of a single specific antigen autoantibody in differentiating cervical cancer patients from normal individuals is relatively low, this invention combines XIRP2 autoantibody, POU3F2 autoantibody, and PCNA autoantibody in parallel to improve diagnostic sensitivity. The results were used to assess the serum of 80 HSIL patients, 32 cervical cancer patients, and 100 normal controls included in the examples, analyzing the value of different specific antigen combinations in cervical cancer diagnosis.

[0094] The method for determining the effectiveness of different specific antigen-autoantibody combinations in the diagnosis of cervical cancer is as follows: When two or three specific antigen-autoantibodies are used in parallel to differentiate cervical cancer, if the result of any one specific antigen-autoantibody in the combination is positive, then the diagnostic result of that specific antigen-autoantibody combination is positive. ROC curves are plotted based on the results, and sensitivity and specificity are statistically analyzed. The results are as follows: Figure 5 As shown in Table 2.

[0095] Table 2 shows the statistical results (N (%)) of different specific antigen-autoantibody combinations used in the diagnosis of cervical cancer.

[0096]

[0097] Note: n represents the total number of samples, N represents the number of antibody-positive samples, and % represents the antibody positivity rate.

[0098] Depend on Figure 5 The results show that the AUC of XIRP2 autoantibody in differentiating cervical cancer patients from normal individuals is 0.76; the AUC of POU3F2 autoantibody is 0.74; and the AUC of PCNA autoantibody is 0.69. The AUC of PCNA+POU3F2 combined in differentiating cervical cancer patients from normal individuals is 0.81; the AUC of XIRP2+PCNA combined in differentiating cervical cancer patients from normal individuals is 0.84; the AUC of XIRP2+POU3F2 combined in differentiating cervical cancer patients from normal individuals is 0.85; and the AUC of XIRP2+PCNA+POU3F2 combined in differentiating cervical cancer patients from normal individuals is 0.92. Therefore, the AUC of the three autoantibodies against tumor-associated antigens in distinguishing cervical cancer patients is greater than 0.5. Thus, XIRP2 autoantibody, POU3F2 autoantibody, and PCNA autoantibody can be used as an adjunct to the diagnosis of cervical cancer.

[0099] Depend on Figure 5As can be seen from Table 1, compared with a single specific antigen autoantibody, the AUC and sensitivity of the combination of multiple specific antigen autoantibodies for the diagnosis of cervical canceration are obviously improved; when three specific antigen autoantibodies (XIRP2+POU3F2+PCNA) are combined, the AUC of the ROC curve reaches a maximum of 0.92, the sensitivity of the detection reaches 42.9%, and has a higher diagnostic value, while the specificity of the detection can still reach 93.0%. Therefore, when multiple specific antigen autoantibodies are combined for the diagnosis of cervical canceration, the diagnostic sensitivity can be greatly improved under the premise of ensuring the diagnostic specificity. In addition, with the increase of the number of specific antigen autoantibodies in the combination, the Youden index continuously increases and gradually tends to 1, indicating that the method of using multiple specific antigen autoantibodies for the diagnosis of cervical canceration has a good diagnostic value.

[0100] The above experimental results show that the ELISA kit of the present application has a higher diagnostic value for cervical canceration, further proving that the ELISA kit of the present application is an ideal early diagnosis and screening method for cervical canceration.

Claims

1. Use of a reagent for detecting a biomarker in the manufacture of a product for screening of cervical carcinogenesis, characterized in that, The biomarker is at least one of XIRP2 autoantibody, POU3F2 autoantibody, PCNA autoantibody.

2. Use according to claim 1, characterized in that, The cervical cancer is cervical intraepithelial neoplasia HSIL II / III or cervical cancer.

3. Use according to claim 1, characterized in that, The reagent is a reagent for detecting the biomarker in a sample by enzyme-linked immunosorbent assay, protein chip, immunoblotting or microfluidic immunoassay.

4. Use according to claim 3, characterized in that, The reagent is an antigen or antibody for detecting the biomarker.

5. Use according to claim 4, characterized in that, The antigen is at least one of XIRP2 protein, POU3F2 protein, PCNA protein.

6. Use according to claim 3, characterized in that, The sample is serum, plasma, interstitial fluid or cervical exfoliated cell sample.

7. Use according to any one of claims 1 to 6, characterized in that, The product is a protein chip, a kit or a preparation.

8. A kit for screening of cervical carcinogenesis, characterized by, The kit comprises a reagent for detecting a biomarker, which is at least one of XIRP2 autoantibody, POU3F2 autoantibody, PCNA autoantibody.

9. The kit of claim 8, wherein The kit detects the biomarker in a sample by enzyme-linked immunosorbent assay, protein chip, immunoblotting or microfluidic immunoassay.

10. The kit of claim 9, wherein The kit is an ELISA detection kit, which comprises a solid phase carrier and an antigen coated on the solid phase carrier; the antigen is at least one of XIRP2 protein, POU3F2 protein, PCNA protein.

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