TS Triage of HR-HPV Positive Population and Its Accuracy Study

A Study on the Application of TS for Triage in HR-HPV Viral Load Positive Population and Accuracy Related Studies

A prospective study assessing TS accuracy in HR-HPV+ women, influencing factors, and 1-year follow-up of false-positives versus biopsy.

Study Overview

• Status: Active, not recruiting
• Conditions: Cervical Cancer; Cervical Precancerous Lesions
• Intervention / Treatment: Diagnostic test: Colposcopy and cervical biopsy; Diagnostic test: Follow-up of TS false-positive patients

Detailed Description

The study will be conducted between 2025.01 and 2027.12 and will assess the diagnostic accuracy of TS testing in patients with positive HR-HPV viral loads by means of a prospective controlled study analyzing the factors influencing the false positive/negative results (age/gynecological findings/vaginal microbiota, etc.) and using cervical biopsy as a reference standard for 1 year in cases of TS false positives. The principal investigator will write and publish a paper.

• Study Type: Observational
• Enrollment (Estimated): 300

Contacts and Locations

Study Locations

• China
  • Jiangsu
    • Nantong, Jiangsu, China, 226001
      • Affiliated Hospital of Nantong University

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Child; Adult; Older Adult
• Accepts Healthy Volunteers: Yes

• Sampling Method: Probability Sample

Study Population

Patients who visited the gynecological outpatient department of the Affiliated Hospital of Nantong University between October 2024 and October 2026, and were detected as positive for high-risk human papillomavirus (HR-HPV) in cervical cancer screening through the Hybrid Capture 2 (HC2) technique, and had indications for colposcopic evaluation, and were scheduled for colposcopy or cervical pathological biopsy, as well as those patients who had both the HPV viral load test results and the Triple Staining (TS) results in our hospital and were awaiting colposcopy, will be selected.

Description

Inclusion Criteria:

1. Patients in our outpatient clinic who have already undergone HPV viral load testing and are awaiting colposcopy, or those who have both HPV viral load test results and TS (Triple Staining) results and are awaiting colposcopy；
2. Informed consent obtained from voluntary participants.

Exclusion Criteria:

1. Patients who are unable to cooperate or have poor compliance;
2. Within four months postpartum;
3. During the first three days of menstruation with heavy flow;
4. Within three months after cervical surgery (including cervical biopsy);
5. Previous radiotherapy in the pelvic region;
6. Currently undergoing chemotherapy or within five weeks after chemotherapy;
7. Clinically evident acute or subacute cervical/vaginal infection;
8. History of photosensitivity disorders, current photodynamic therapy, or exposure to photosensitizing agents;
9. Suspected or confirmed history of alcohol/drug abuse, or any other condition that-in the investigator's judgment-may reduce enrollment feasibility or complicate participation (e.g., frequent job relocation, unstable living conditions leading to high risk of loss to follow-up);
10. Other patients deemed ineligible by the investigators.

Study Plan

How is the study designed?

Number of groups / cohorts

6

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
HPV16/18 - Low VL; HPV type 16/18 positive, low viral load group (1.00-99.99 RLU/PC)	Diagnostic test: Colposcopy and cervical biopsy; The cervix was visualized through colposcopic magnification and samples of suspicious tissue were taken for pathological examination.; Diagnostic test: Follow-up of TS false-positive patients; False-positive people with initial TS screening were followed up for 1 year, with a second TS examination six months after the initial TS screening.
HPV16/18 - Medium VL; HPV type 16/18 positive, medium viral load group (100-999.99 RLU/PC)	Diagnostic test: Colposcopy and cervical biopsy; The cervix was visualized through colposcopic magnification and samples of suspicious tissue were taken for pathological examination.; Diagnostic test: Follow-up of TS false-positive patients; False-positive people with initial TS screening were followed up for 1 year, with a second TS examination six months after the initial TS screening.
HPV16/18 - High VL; HPV type 16/18 positive, high viral load group (≥1000.00 RLU/PC)	Diagnostic test: Colposcopy and cervical biopsy; The cervix was visualized through colposcopic magnification and samples of suspicious tissue were taken for pathological examination.; Diagnostic test: Follow-up of TS false-positive patients; False-positive people with initial TS screening were followed up for 1 year, with a second TS examination six months after the initial TS screening.
Other12 HR-HPV - Low VL; Positive for 12 other high-risk HPV types, low viral load group (1.00-99.99 RLU/PC)	Diagnostic test: Colposcopy and cervical biopsy; The cervix was visualized through colposcopic magnification and samples of suspicious tissue were taken for pathological examination.; Diagnostic test: Follow-up of TS false-positive patients; False-positive people with initial TS screening were followed up for 1 year, with a second TS examination six months after the initial TS screening.
Other12 HR-HPV - Medium VL; Positive for 12 other high-risk HPV types, medium viral load group (100-999.99 RLU/PC)	Diagnostic test: Colposcopy and cervical biopsy; The cervix was visualized through colposcopic magnification and samples of suspicious tissue were taken for pathological examination.; Diagnostic test: Follow-up of TS false-positive patients; False-positive people with initial TS screening were followed up for 1 year, with a second TS examination six months after the initial TS screening.
Other12 HR-HPV - High VL; Positive for 12 other high-risk HPV types, high viral load group (≥1000.00 RLU/PC)	Diagnostic test: Colposcopy and cervical biopsy; The cervix was visualized through colposcopic magnification and samples of suspicious tissue were taken for pathological examination.; Diagnostic test: Follow-up of TS false-positive patients; False-positive people with initial TS screening were followed up for 1 year, with a second TS examination six months after the initial TS screening.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Sensitivity of TS for diagnosing populations with different HR-HPV subtypes and viral loads	Using cervical biopsy pathology as the gold standard, the study participants were stratified into 6 detailed subgroups based on HR-HPV genotype (HPV 16/18 and other 12 genotypes) and viral load gradients (low: 1.00-99.99; medium: 100-999.99; high: ≥1000.00) to form six detailed subgroups. The percentage of TS-positive cases among pathologically confirmed patients was calculated for each subgroup to determine the differences in diagnostic sensitivity when using TS as a screening tool.	January 2025 to December 2025
The specificity of TS in diagnosing individuals with different HR-HPV subtypes and viral loads	Using the pathological diagnosis of cervical biopsy as the gold standard, the study subjects were stratified into 6 detailed subgroups based on HR-HPV genotype (HPV 16/18 and other 12 genotypes) and viral load gradients (low: 1.00-99.99; medium: 100-999.99; high: ≥1000.00) to form six detailed subgroups. The percentage of TS-negative cases among pathologically negative (non-lesional) patients was calculated for each subgroup to clarify the differences in clinical efficacy of the test as a triage tool in excluding non-lesional individuals and reducing unnecessary referral rates.	January 2025 to December 2025
The positive predictive value of TS in populations with different HR-HPV subtypes and viral loads	Using cervical biopsy pathology as the gold standard, the study participants were stratified into 6 detailed subgroups based on HR-HPV genotype (HPV 16/18 and other 12 genotypes) and viral load gradients (low: 1.00-99.99; medium: 100-999.99; high: ≥1000.00) to form six detailed subgroups. The percentage of TS-positive subjects in each subgroup who were pathologically confirmed to have lesions was calculated to assess the predictive ability of TS-positive results for cervical high-grade lesions and their clinical diagnostic value across different viral load levels.	January 2025 to December 2025
Negative predictive value of TS in populations with different HR-HPV subtypes and viral loads	Using cervical biopsy pathology as the gold standard, the study participants were stratified into 6 detailed subgroups based on HR-HPV genotype (HPV 16/18 and other 12 genotypes) and viral load gradients (low: 1.00-99.99; medium: 100-999.99; high: ≥1000.00) to form six detailed subgroups. The percentage of subjects with a normal/non-lesional pathological diagnosis among those with a negative TS test result was calculated for each subgroup to assess the reliability of TS-negative results in ruling out cervical lesions and to determine the safety of using TS as a triage tool at different viral load levels.	January 2025 to December 2025
The AUC of TS in diagnosing individuals with different HR-HPV subtypes and viral loads	Using the pathological diagnosis of cervical biopsy as the gold standard, the study subjects were stratified into 6 detailed subgroups based on HR-HPV genotyping (HPV types 16/18 and other 12 types) and viral load gradients (low: 1.00-99.99; medium: 100-999.99; high: ≥1000.00) to form six detailed subgroups. By plotting receiver operating characteristic (ROC) curves and calculating the area under the curve (AUC), we comprehensively evaluated the overall diagnostic performance of TS in distinguishing between women with and without cervical lesions across different viral load levels. The closer the AUC value is to 1.0, the higher the accuracy of TS in classifying patients within that subgroup.	January 2025 to December 2025

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Factors Contributing to False-Positive Results in TS	This study used pathological diagnosis based on cervical biopsy as the gold standard. Patients with positive TS test results but pathological diagnoses of normal or inflammation were defined as the TS false-positive group, while the TS true-positive group-in which TS and pathological results were consistent-served as the control group. Univariate and multivariate logistic regression analyses were performed on the two groups, examining factors such as age, cervical conditions (e.g., columnar epithelium ectopia, hypertrophy, polyps), history of cervical conization, and vaginal microbiome (various types of vaginal inflammation and pathogen infections), with the aim of identifying independent risk factors contributing to false-positive interference in TS results.	January 2025 to December 2025
Factors Contributing to False-Negative Results in TS	This study used the pathological diagnosis of cervical biopsy as the gold standard, defining patients with negative TS test results but a pathological diagnosis of lesions as the TS false-negative group, and using the TS true-negative group-in which TS and pathological results were consistent-as the control group. Through univariate and multivariate logistic regression analyses of indicators such as age, cervical anatomical status, and vaginal microbiome composition in both patient groups, the study aims to identify independent risk factors leading to TS missed diagnosis (false negatives) and provide clinical evidence to optimize screening strategies.	January 2025 to December 2025
Sensitivity of TS in predicting 1-year follow-up outcomes in patients with a positive initial screening result but a negative cervical biopsy	This indicator uses the pathological diagnoses from the second and third cervical biopsies during the 1-year follow-up period as the gold standard. For the specific population of patients who were "TS-positive but biopsy-negative" at the initial visit, it calculates the percentage of patients in this group who tested positive on subsequent TS screening during the follow-up period out of the total number of patients in this group who were ultimately pathologically confirmed to have lesions. This indicator is designed to evaluate the ability of TS to detect missed diagnoses at the initial visit, occult lesions, or newly developed lesions during long-term monitoring, to clarify its safety as a follow-up management tool, and to prevent the missed diagnosis of high-grade lesions.	Baseline (initial visit), 6-month follow-up, 12-month follow-up.
Specificity of TS in predicting 1-year follow-up outcomes for patients with a positive initial screening result but a negative cervical biopsy	This indicator uses the pathological diagnoses from the second and third cervical biopsies during a 1-year follow-up period as the gold standard. For a specific subgroup of patients who were "TS-positive but biopsy-negative" at their initial visit, it calculates the percentage of patients in this subgroup who maintained a consistently negative TS result during follow-up, relative to the total number of patients in this subgroup who ultimately maintained a normal or inflammatory pathological diagnosis. This indicator aims to evaluate the ability of TS to rule out transient risks and identify non-lesional cases, thereby demonstrating its clinical efficacy in reducing unnecessary repeat biopsies and optimizing the allocation of medical resources during the follow-up process.	Baseline (initial visit), 6-month follow-up, 12-month follow-up.
The positive predictive value of TS for predicting outcomes at 1-year follow-up in patients with a positive initial screening result but a negative cervical biopsy	This indicator uses the pathological diagnoses from the second and third cervical biopsies during the 1-year follow-up period as the gold standard. For the specific subgroup of patients who were "TS-positive but biopsy-negative" at the initial visit, it calculates the percentage of patients among all those with positive TS results during follow-up who were ultimately pathologically confirmed to have lesions. This indicator aims to evaluate the predictive value of TS "false-positive" results in identifying actual lesions during follow-up, thereby providing data to support clinical decisions regarding the timing of intervention.	Baseline (initial visit), 6-month follow-up, 12-month follow-up.
The negative predictive value of TS for predicting outcomes at 1-year follow-up in patients with a positive initial screening result but a negative cervical biopsy	This indicator uses the pathological diagnoses from the second and third cervical biopsies during the 1-year follow-up period as the gold standard. For the specific subgroup of patients who were "TS-positive but biopsy-negative" at the initial visit, it calculates the percentage of patients who maintained a normal/negative pathological result throughout the follow-up period among those with negative TS results on all follow-up tests. This indicator is used to evaluate the safety of TS-negative results in follow-up management, providing a basis for using it as a decision-making tool to extend follow-up intervals and reduce monitoring frequency.	Baseline (initial visit), 6-month follow-up, 12-month follow-up.
The AUC of TS for predicting 1-year follow-up outcomes in patients with a positive initial screening result but a negative cervical biopsy	This metric uses the pathological diagnoses from the second and third cervical biopsies during the 1-year follow-up period as the gold standard. For the specific subgroup of patients who were "TS-positive but biopsy-negative" at the initial visit, a ROC curve is plotted and the area under the curve (AUC) is calculated by integrating dynamic testing data collected over the 1-year follow-up period. This indicator aims to comprehensively evaluate the overall predictive accuracy of TS regarding long-term disease outcomes in this population and to compare the monitoring efficacy of TS alone versus TS combined with HPV testing.	Baseline (initial visit), 6-month follow-up, 12-month follow-up.

Collaborators and Investigators

• Sponsor: Affiliated Hospital of Nantong University
• Investigators: Principal Investigator: Yuquan Zhang Professor, Affiliated Hospital of Nantong University

Study record dates

Study Major Dates

• Study Start (Actual): October 1, 2024
• Primary Completion (Estimated): December 31, 2027
• Study Completion (Estimated): December 31, 2027

Study Registration Dates

• First Submitted: August 17, 2026
• First Submitted That Met QC Criteria: August 17, 2026
• First Posted (Actual): August 20, 2026

Study Record Updates

• Last Update Posted (Actual): August 20, 2026
• Last Update Submitted That Met QC Criteria: August 17, 2026
• Last Verified: August 1, 2026

More Information

Terms related to this study

• Additional Relevant MeSH Terms: Urogenital Diseases; Genital Diseases; Urogenital Neoplasms; Neoplasms by Site; Neoplasms; Female Urogenital Diseases; Female Urogenital Diseases and Pregnancy Complications; Uterine Diseases; Genital Diseases, Female; Genital Neoplasms, Female; Uterine Cervical Diseases; Uterine Neoplasms; Uterine Cervical Neoplasms; Diagnostic Techniques and Procedures; Diagnosis; Surgical Procedures, Operative; Minimally Invasive Surgical Procedures; Diagnostic Techniques, Surgical; Endoscopy; Urogenital Surgical Procedures; Gynecologic Surgical Procedures; Obstetric Surgical Procedures; Diagnostic Techniques, Obstetrical and Gynecological; Colposcopy
• Other Study ID Numbers: Tdfy-FCK-2025

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No