Cervical lesion risk prediction model based on combination of double-gene methylation and clinical characteristics and application thereof

Abstract

The invention relates to the technical field of cervical lesions, and discloses a cervical lesion risk prediction model based on combination of double gene methylation and clinical characteristics and application thereof, and the method comprises the following steps: S1, selecting a research object; s2, carrying out double-gene methylation detection; and S3, data statistical analysis: performing statistical analysis by applying SPSS 25.0 and R4.1. 2 software, performing chi-square test and single-factor logistic regression analysis on each potential influence factor, and bringing the screened influence factors into multi-factor logistic regression analysis. According to the cervical lesion risk prediction model based on combination of double-gene methylation and clinical characteristics and application and development thereof, evidence-based basis can be better provided, clinical reduction of missed diagnosis and misdiagnosis rates of cervical cancer is helped, morbidity and mortality are reduced, and implementation of a strategic target of eliminating cervical cancer in China is assisted.

Description

TECHNICAL FIELD

[0001] The application relates to the technical field of cervical lesions, in particular to a cervical lesion risk prediction model based on double-gene methylation combined with clinical characteristics and application thereof. BACKGROUND

[0002] Cervical cancer, also known as cervical carcinoma, is a malignant tumor occurring at the cervical part of a female. It is one of the main causes of female cancer morbidity and mortality worldwide, and has a high morbidity and mortality.

[0003] Methylation detection is expensive, and although methylation detection is mainly used as a shunting means based on abnormal cervical cancer screening in current research, multi-gene detection is expensive and difficult to be widely promoted in clinical practice. SUMMARY

[0004] The purpose of the present application is to provide a cervical lesion risk prediction model based on double-gene methylation combined with clinical characteristics and application thereof, so as to solve the problems raised in the background.

[0005] To achieve the above-mentioned purpose, the present application provides the following technical scheme: a cervical lesion risk prediction model based on double-gene methylation combined with clinical characteristics, comprising the following steps: S1, selecting research subjects: in this study, patients who visited the gynecological outpatient department of the hospital and 15 Qin-Ba mountain area cooperative hospitals from January 2025 to June 2026 and met part of the application scenarios and inclusion criteria were selected as research subjects; S2, double-gene methylation detection: patients who need to be referred to colposcopy according to the current domestic guidelines were sampled for double-gene methylation detection before colposcopy; S3, data statistical analysis: statistical analysis was performed by using SPSS 25.0 and R4.1.2 software, chi-square test and single-factor logistic regression analysis were performed on each potential influencing factor, and the screened influencing factors were included in the multi-factor logistic regression analysis.

[0006] Preferably, the application scenarios of double-gene methylation detection in S1 are as follows: (1) stratified management of high-risk HPV positive or cytological abnormal women; (2) risk assessment of cervical Ⅲ transformation zone and potential adenocarcinoma patients; (3) monitoring after cervical lesion treatment; (4) evaluation of withdrawal from cervical cancer screening plan.

[0007] In addition to meeting the application scenarios, the following inclusion criteria must be met: (1) all patients received TCT and HPV detection; (2) no sexual life, no vaginal irrigation and vaginal medication within 3 days; (3) No anti-human papillomavirus treatment in the past 6 months; (4) All patients come from counties in the Qin-Ba mountainous area; (5) Patients who cooperate with the examination, have complete data and complete the informed consent form.

[0008] Exclusion criteria when meeting the inclusion criteria: (1) menstruating women; (2) Known to have other malignant tumors other than cervical lesions (cancer); Unable to complete follow-up examination and follow-up; (4) History of autoimmune disease or taking immunosuppressive agents; (5) Pregnant or lactating women.

[0009] Data collection of patients meeting the above conditions: (1) General information [outpatient number, name, age, height, weight, body mass index, whether menopausal]; Cervical liquid-based cytology results (TCT); HPV typing results; Cervical transformation zone type under colposcopy; Cervical histopathology; Vaginal microecological conditions.

[0010] Age, body mass index, menopausal status, cervical liquid-based cytology results, HPV typing results, cervical transformation zone type under colposcopy, and vaginal microecological conditions are used as influencing factors in the prediction model.

[0011] Preferably, the human PAX1 and JAM3 double gene methylation detection kit used in S2 is provided by Beijing Origin Polymer Biotechnology Co., Ltd.

[0012] Preferably, if biopsy is needed under colposcopy in S2, biopsy should be taken for pathological diagnosis, and pathology is the final diagnostic gold standard.

[0013] The application also provides an application of a cervical lesion risk prediction model based on double gene methylation combined with clinical characteristics, which helps to improve the HPV-related epidemiological database in the Qin-Ba mountainous area, optimize the cervical screening strategy, realize the diversion management of patients, detect high-risk groups in time, reduce the harm caused by excessive diagnosis and treatment, and provide help for clinical treatment in risk assessment, guidance, prognosis evaluation and recurrence detection.

[0014] Compared with the prior art, the application has the following beneficial effects: The cervical lesion risk prediction model based on double gene methylation combined with clinical features and its application can reveal the HPV type distribution characteristics in the Qinba mountainous area, and provide targeted theoretical guidance for cervical cancer vaccination and cervical lesion prevention and treatment in the region, thereby providing data support for domestic vaccine research and development, breaking the monopoly situation of foreign enterprises, and having a promising future. The establishment of the cervical lesion prediction model can alleviate patient anxiety and over-treatment, reduce medical costs, and avoid waste of medical resources. Early diagnosis and treatment of cervical cancer can significantly reduce the financial pressure of gynecological tumor treatment in the region and prevent high costs generated by cervical cancer treatment.

[0015] The cervical lesion risk prediction model based on double gene methylation combined with clinical features and its application further perfects the HPV-related epidemiological database in the Qinba mountainous area, innovatively optimizes the cervical screening strategy, constructs the cervical lesion risk prediction model, realizes the diversion management of patients, detects high-risk groups in time while reducing the harm caused by excessive diagnosis and treatment, and can provide help for clinical treatment in the scenes of risk assessment, guidance, prognosis evaluation, and recurrence detection.

[0016] The cervical lesion risk prediction model based on double gene methylation combined with clinical features and its application will help to better provide evidence-based basis, help clinical doctors to reduce the misdiagnosis rate of cervical cancer, reduce the incidence and mortality, and help to realize the strategic goal of eliminating cervical cancer in China. BRIEF DESCRIPTION OF DRAWINGS

[0017] Fig. 1 The double gene methylation screening technology roadmap for cervical cancer is constructed. Fig. 2 The double gene methylation prediction model construction technology roadmap is constructed. DETAILED DESCRIPTION

[0018] The technical solutions in the embodiments of the present application will be described clearly and completely in combination with the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor are within the scope of protection of the present application.

[0019] Please refer to Figs. 1-2 The present application provides the following technical solutions: A cervical lesion risk prediction model based on double gene methylation combined with clinical features and its application, comprising the following steps: S1, Take the research object: This study takes the patients who visited the gynecological clinic of our hospital and 15 cooperative hospitals in Qinba mountain area from January 2025 to June 2026 and met the part of the application scenario and the inclusion criteria as the research object; The application scenario of double gene methylation detection: (1) stratified management of high-risk HPV positive or cytological abnormal women; (2) risk assessment of cervical Ⅲ transformation zone and potential adenocarcinoma patients; (3) monitoring after treatment of cervical lesions; (4) evaluation of patients who have withdrawn from the cervical cancer screening program.

[0020] In addition to meeting the application scenario, the following inclusion criteria must be met: (1) all patients received TCT and HPV detection; (2) no sexual intercourse, no vaginal irrigation and vaginal medication within 3 days; (3) no anti-human papillomavirus treatment within 6 months; (4) all patients come from counties in Qinba mountain area; (5) patients who cooperate with the examination, have complete data and complete the informed consent form; Exclusion criteria when meeting the inclusion criteria: (1) menstruating women; Patients with known cervical lesions (cancer) and other malignant tumors; Patients who cannot complete follow-up examination and follow-up; (4) patients with a history of autoimmune disease or taking immunosuppressive agents; (5) pregnant or lactating women.

[0021] Data collection of patients who meet the above conditions: (1) general information; Cervical liquid-based cytology results (TCT); HPV typing results; Cervical transformation zone type under colposcopy; Cervical histopathology; Vaginal microecology.

[0022] Age, body mass index, menopause or not, cervical liquid-based cytology results, HPV typing results, cervical transformation zone type under colposcopy, and vaginal microecology were included in the prediction model as influencing factors.

[0023] S2, Double gene methylation detection: patients who need to be referred to colposcopy according to the current domestic guidelines were sampled for double gene methylation detection before colposcopy. Vaginal microecology examination was performed before methylation detection. PAX1 and JAM3 double gene methylation detection kits were used in double gene methylation detection, which were provided by Beijing Origin Polymer Biotechnology Co., Ltd.

[0024] Colposcopy and cervical biopsy: The patient was placed in lithotomy position, and colposcopy was performed. The surface mucus of the cervix was cleaned with normal saline, and the natural state of the cervix was observed, including the shape and color. A wet cotton ball coated with 3%-5% acetic acid was applied to the cervix, and after 120 seconds, the cotton ball was removed. The color change of the cervix was observed directly, and the type of transformation zone, acetowhite reaction, mosaic blood vessels, and atypical blood vessels were observed. The presence, partial presence, and absence of squamous-columnar junction were corresponding to type I, type II, and type III transformation zone, respectively. The suspicious lesion area was taken by biopsy forceps under colposcopy, and the sample was fixed with 10% formaldehyde. The cervical tissue was immediately sent to the pathology department for sectioning after fixation. Hematoxylin-eosin staining was used, and the results were provided by the pathologist.

[0025] S3, SPSS 25.0 and R4.1.2 software were used for statistical analysis. The potential influencing factors were subjected to chi-square test and single-factor logistic regression analysis. The selected influencing factors were included in the multi-factor logistic regression analysis.

[0026] During use, the methylation detection in our hospital and 15 Qin-Ba mountain area cooperative hospitals was completed in our hospital precision diagnosis center, and the detection report was issued. Cervical sampling swabs were used to collect cervical exfoliative cells, which were stored in sample tubes containing cell preservation solution and stored and transported at room temperature.

[0027] The label indicated the patient's name, age, date, and sampling hospital. The detection report was issued within 7 days after sampling. After 2-5 ml of cervical exfoliative cell specimens were collected for DNA extraction, DNA concentration and quality determination were performed, and 20 ul of DNA was used for bisulfite conversion.

[0028] After bisulfite conversion, DNA was detected by real-time fluorescence quantitative PCR (real-time PCR) using a real-time fluorescence quantitative PCR instrument according to the human PAX1 and JAM3 gene methylation detection kit instructions. The sample target gene ΔCt value was recorded, JAM3 result was represented by ΔCtJ, >10 was negative, PAX1 result was represented by ΔCtP, >6.0 was negative, and the lower the ΔCt value, the higher the methylation degree of the target gene.

[0029] Positive clinical significance suggested high lesions (cancer) and recommended colposcopy for careful examination of the cervix or vaginal wall; or close follow-up; or CIN2 / 3 progression probability was high, and further referral for colposcopy or pathological examination was recommended.

[0030] Negative clinical significance at the same time suggested 6-12 months of follow-up, or high conversion rate or suggested possible conversion.

[0031] The data of 6-12 months follow-up were statistically analyzed by SPSS25.0 and R4.1.2 software. The potential influencing factors were subjected to chi-square test and single factor logistic regression analysis. The screened influencing factors were included in the multivariate logistic regression analysis.

[0032] The independent influencing factors were introduced into R4.1.2 to establish a nomogram model for individual prediction of CIN2+ lesion risk.

[0033] The internal validation of the nomogram model applied Bootstrap self-sampling method. The discrimination and calibration of the nomogram model were evaluated by the consistency index (C-index), the area under the receiver operating characteristic curve (AUC), and the calibration curve. The clinical benefit of the nomogram model was assessed by the decision curve.

[0034] When AUC>0.7, the predictive value of the model was better.

[0035] The goodness of fit of the model was evaluated by Hosmer Lemeshow test. If the P value of the test was >0.05, the model was considered to have good goodness of fit.

[0036] The AUC of the ROC curve and its 95% confidence interval were calculated. The probability value corresponding to the maximum Youden index was selected as the optimal cut-off value of the model.

[0037] The sensitivity, specificity, accuracy, AUC, and Youden index of the nomogram model and each individual test were calculated. McNemar test and Z test were used for comparison.

[0038] The difference of bilateral test was taken as the test level of α=0.05.

[0039] The research results were popularized in primary medical institutions in Qinba mountainous area. The cooperative units conducted relevant lectures or training. The synergistic effect of medical association was actively played. The disease control agencies and community health service centers jointly constructed the immune prevention barrier of cervical cancer, which could provide data and theoretical support for disease control vaccination consultation clinics, cervical / gynecological clinics, and better provide HPV-related consultation services, health education, and popular science propaganda for patients, and improve the medical service ability of comprehensive prevention and control of cervical cancer in the region.

[0040] Embodiments of the present application are described, which, while including the best mode known to the inventors to date of carrying out the application, are provided for illustrative purposes only. It is understood that various modifications, substitutions, changes, and alterations to the embodiments can be made by one of ordinary skill in the art without departing from the spirit and scope of the present application, which is defined by the following claims and their equivalents.

Claims

1. A cervical lesion risk prediction model based on double gene methylation combined with clinical features, characterized by: Prepared by the following steps: S1, select the research object: this study from January 2025 to June 2026 in our hospital and 15 Qinba mountainous area of gynecological clinic and at the same time meet part of the application scene and inclusion criteria for patients as the research object; S2, double gene methylation detection: patients according to the current domestic guidelines need to be referred to colposcopy, before colposcopy sampling for double gene methylation detection; S3, data statistical analysis: SPSS 25.0 and R4.1.2 software were used for statistical analysis, and the potential influencing factors were subjected to chi-square test and single factor logistic regression analysis. The selected influencing factors were included in the multivariate logistic regression analysis.

2. The cervical lesion risk prediction model based on double gene methylation combined with clinical characteristics according to claim 1, characterized in that: The S1 application scene of double gene methylation detection: (1) stratified management of high-risk HPV positive or cytological abnormal women; (2) risk assessment of cervical Ⅲ transformation zone and potential adenocarcinoma patients; (3) monitoring after treatment of cervical lesions; (4) evaluation of withdrawal from cervical cancer screening program; In line with the application scene and the following inclusion criteria: (1) all received TCT, HPV detection; (2) no sexual life, no vaginal irrigation and vaginal medication within 3 days; (3) no anti-human papillomavirus treatment within 6 months; (4) patients from Qinba mountainous area counties; (5) cooperate with the examination, complete data and complete the informed consent form; Exclusion criteria when meeting the inclusion criteria: (1) menstrual period; (2) known to have other malignant tumors except cervical lesions; Unable to complete follow-up examination and follow-up; (4) have a history of autoimmune disease, or are taking immunosuppressive agents; (5) pregnant or lactating women; Data collection of patients meeting the above conditions: (1) general information; Cervical liquid-based cytology results; HPV typing results; Cervical transformation zone type under colposcopy; Cervical histopathology; (6) vaginal microecological conditions.

3. The cervical lesion risk prediction model based on double gene methylation combined with clinical characteristics according to claim 2, characterized in that: The double gene methylation detection in S2 uses human PAX1 and JAM3 double gene methylation detection kit, which is provided by Beijing Origin Polymer Biotechnology Co., Ltd.

4. The cervical lesion risk prediction model based on double-gene methylation combined with clinical characteristics according to claim 3, characterized in that: If biopsy is needed under colposcopy in S2, biopsy should be taken for pathological diagnosis, and pathology is the final diagnosis gold standard.

5. Use of a cervical lesion risk prediction model based on dual-gene methylation in combination with clinical features, characterized in that: The cervical lesion risk prediction model based on double gene methylation combined with clinical characteristics as claimed in any one of claims 1-4 can be popularized in primary medical institutions in Qinba mountainous area, providing data and theoretical support for disease control, vaccine inoculation consultation clinics, cervical / gynecological clinics, and improving the medical service ability of cervical cancer comprehensive prevention and control.

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