Wen Chen

HPV testing with 16/18 genotyping for risk stratification among women with normal cytology: a multicenter prospective cohort study from China

ABSTRACT To evaluate the clinical performance of Hybribio’s 14-type HPV real-time PCR with 16/18 genotyping (HBRT-H14) and its risk stratification utility among women with normal cytology (NILM). From 2017 to 2020, a multicenter cohort enrolled 8,401 women aged 30–64 years with NILM cytology. Baseline HPV testing used HBRT-H14. Women positive for HPV 16/18 were referred for colposcopy; follow-up was annual for 3 years or until the detection of cervical intraepithelial neoplasia grade 2 or worse (CIN2+). Analyses included 6,679 women who completed follow-up. Overall HPV positivity was 11.4%, including 2.3% HPV 16/18. Over 3 years, sensitivity and specificity of HPV positivity for CIN2+ were 92.3% (95% confidence interval [CI]: 84.2–96.4) and 89.6% (88.8–90.3). For HPV 16/18 positivity, sensitivity and specificity were 41.0% (30.8–52.1) and 98.2% (97.8–98.5). Three-year cumulative CIN2+ risk was 20.9% (15.2–28.1) for HPV 16/18-positive women, 6.6% (4.9–8.9) for other types, and 0.1% (0.04–0.2) for HPV-negative women. HBRT-H14 shows strong clinical performance for detecting CIN2+, and HPV 16/18 genotyping provides effective risk stratification among women with NILM cytology. Findings support integration of HBRT-H14 into HPV-based screening pathways with HPV 16/18 genotyping and cytology triage of other types. IMPORTANCE This multicenter prospective study evaluated the Hybribio 14 high-risk HPV real-time PCR assay (HBRT-H14) in 8,401 women with normal (NILM) cytology under guideline-based follow-up. The assay showed high clinical sensitivity and a very low risk among HPV-negative women, and HPV 16/18 genotyping provided clear risk stratification. These findings deliver large-scale, practice-oriented evidence supporting integration of HBRT-H14 into HPV-based screening pathways that use HPV 16/18 genotyping with cytology triage of other types.

Application of P16 /Ki‐67 dual‐staining for the detection of high‐grade cervical lesions in the triage of patients with minor abnormal cytology: A meta‐analysis

Abstract Background The p16/Ki‐67 dual‐staining is increasingly applied to increase diagnostic accuracy in detecting high‐grade cervical lesions, including cervical intraepithelial neoplasia Grade 2 (CIN2+) and CIN3+. Objectives To compare the diagnostic performance of p16/Ki‐67 dual‐staining with the human papillomavirus (HPV) tests in the triage of women with atypical squamous cells of undetermined significance (ASC‐US) and low‐grade squamous intraepithelial lesions (LSIL) cytology results. Search Strategy Publications before April 27, 2024, were identified through PubMed, Embase, Web of Science, and Cochrane Library. Selection Criteria The studies have head‐to‐head comparison of p16/Ki‐67 dual‐staining and HPV testing in detecting high‐grade cervical lesions. Data Collection and Analysis Two researchers independently screened articles by title, abstract, and full text. The GRADE and QUADAS‐2 tool was used for quality evaluation. The pooled sensitivity and specificity for CIN2+ and CIN3+ were estimated using random effects models, with results and heterogeneity assessments presented in forest plots. Pretest‐posttest probability (PPP) plots were constructed to evaluate the detection rates of CIN3+ in ASC‐US and LSIL patients. Main Results Twenty‐one studies with 6394 participants were included. The pooled specificity of p16/Ki‐67 dual‐staining was higher than that of HPV tests (CIN2+: 0.73 [95% confidence interval [CI] 0.65–0.80] versus 0.41 [95% CI 0.33–0.50]; CIN3+: 0.61 [95% CI 0.53–0.69] versus 0.33 [95% CI 0.23–0.45]). Summary receiver operating characteristic curve analysis demonstrated p16/Ki‐67 dual‐staining had better diagnostic accuracy for CIN2+ than HPV tests (area under the curve: 0.88 [95% CI 0.85–0.90] versus 0.79 [95% CI 0.75–0.82]). Pretest‐posttest probability (PPP) plots highlighted the superior performance of p16/Ki‐67 dual‐staining for colposcopy referrals in LSIL patients. Conclusions P16/Ki‐67 dual‐staining offers greater specificity for detecting CIN2+/CIN3+ in ASC‐US and LSIL triage, particularly for LSIL. It holds potential as an alternative to HPV tests in resource‐limited settings or LSIL cases, warranting further research to refine its application in diverse populations.

The Distribution of HR‐HPV E6/E7 DNA and mRNA by Histological Grade and the Clinical Performance for Detection of Cervical Cancer and Precancer

ABSTRACT High‐risk human papillomavirus (HR‐HPV) testing, utilizing both DNA and RNA methods, offers enhanced sensitivity compared to cytology for detecting high‐grade cervical intraepithelial neoplasia (CIN2+). Meanwhile, HR‐HPV E6 and E7 mRNAs are more likely to differentiate the transient infection from the persistent than DNA. Aptima HPV can not only detect HPV mRNA but also HPV DNA though it is much more efficient at detecting HPV RNA than DNA. Currently, there are few studies on the distribution of HPV E6 and E7 mRNA and DNA in the same individual. It is interesting to compare the clinical performance of the Onclarity and Aptima HPV assays and to assess variations in viral load across different histological grades at both DNA and mRNA levels. The analysis included 1607 women (902 from a cervical cancer screening population and 705 inpatients and outpatients), with cervical cytological samples tested using the Aptima and Onclarity HPV assays. Both assays demonstrated high agreement for HPV types 18/45 and 16. Signal‐to‐cutoff (S/CO) values and Ct values for various HR‐HPV types increased with histological severity from normal tissue to cancer. Notably, HPV18 Ct values exceeded those for HPV16 and 45 in women with ≥ CIN1 lesions but decreased sharply in cancer cases. Across the screening population, both assays showed similar sensitivity and predictive values for detecting CIN2+ lesions. The area under the curve (AUC) for CIN2+ and CIN3+ detection in the study population was robust (CIN2+: 0.880, 0.863; CIN3+: 0.881, 0.863). The DNA level for various HR‐HPV types increased with histological severity from normal tissue to cancer, which might impact the performance of Aptima HPV assay. Both assays showed similar sensitivity and predictive values for detecting CIN2+ lesions.

Association of HPV16 /18 genotype infection with the Silva pattern classification system in human papilloma virus‐associated endocervical adenocarcinomas

Abstract Objective Persistent high‐risk human papillomavirus (HR‐HPV) infection is an essential risk factor for HPV‐associated adenocarcinomas (HPVA). A three‐tier pattern system (the Silva pattern) for endocervical adenocarcinoma (ECA) associated with tumor metastasis and recurrence was described by Elvio G. Silva nearly 10 years ago. However, there are no studies on the association between HPV genotypes and Silva patterns. Methods The Silva pattern classification was performed on 240 surgical HPVA specimens according to the 2020 World Health Organization classification of female genital tract cancers. HPV DNA was detected using the SPF 10 ‐DEIA‐LiPA 25 assay for all specimens and an attribution algorithm was used to calculate the attribution rate of HPV16/18. Results Out of all HPVA cases, 29 patients (12.1%) were found to have tumors with Silva pattern A, 122 (50.8%) had pattern B tumors, and 89 (37.1%) had pattern C (representing the worst morphological behavior, poorest prognosis and highest risk of mortality). The crude prevalence of HPV16 and 18 was 46.9% and 44.7%, respectively. The attribution of HPV16 in Silva patterns A, B, and C was 58.0%, 51.7%, and 33.8%, respectively ( P  = 0.123). Similarly, the attribution of HPV18 was found to be 29.8%, 39.7%, and 49.5% in patterns A, B, and C, respectively. Notably, there was a statistically significant positive linear relationship between the prevalence of HPV18 and the Silva pattern from A to C ( P  = 0.002). Conclusion HPV16 and 18 are the most prevalent HPV subtypes in patients with HPVA. HPVA patients who are infected with HPV18 exhibit worse morphological behavior compared to those with the HPV16 genotype.

The variations in the natural history of high‐risk human papillomavirus infections in Chinese healthy women aged 27–45 years compared with 18–26 years: A prospective cohort study

AbstractData investigating the natural history of high‐risk human papillomavirus (HR‐HPV) infection in mid‐adult women compared with young adult women from regions exhibiting a bimodal distribution pattern are scarce. From November 2012 to September 2019, 3681 healthy women aged 18–45 years from the control group of a bivalent HPV vaccine Phase 3 trial in China were followed over 5.5 years. At scheduled visits (Day 0, months 7, 12, 18, 24, 30, 42, 54, and 66), cervical samples were collected for ThinPrep Pap tests and HPV DNA testing, women with abnormal cytology were referred for colposcopy. Data was analyzed using Cox regression model and a competing risk model. Sensitivity analyses were performed among participants attending all scheduled visits. The incidences of HR‐HPV persistent infections (over 6 months [6mPIs]) were 35.5 and 29.0 per 1000 person‐years (PYs) (hazard ratio [HR] = 1.21, 95% confidence interval [CI]: 1.00, 1.46), and HR‐HPV associated CIN grade 2 or greater (CIN2+) were 4.3 and 1.9 per 1000 PYs (HR = 2.31, 95% CI: 1.25, 4.26) in women aged 18–26 and 27–45 years. Competing risk models showed that the cumulative incidence of HR‐HPV infections that progressed to CIN2+ was significantly higher in women aged 18–26 than in women aged 27–45 (5.3% vs. 2.9%, Gray's test p = .0291). The cumulative clearance rates of HR‐HPV infections in women aged 18–26 and 27–45 were similar (94.7% vs. 95.8%, Gray's test p = .3309) during the study period. In conclusion, although mid‐adult women exhibit lower incidences of HR‐HPV infection and associated cervical lesions compared to young women, this population continues to face a substantial risk of acquiring causal HPV infections, which may progress to cervical lesion.

Concordance between the BD Onclarity and Roche cobas assays for detection of HPV DNA in a Chinese population

AbstractAs cervical cancer screening shifts from cytology to human papillomavirus (HPV) testing, a major issue involves validating more HPV tests. In recent years, some HPV tests are used for clinical performance verification in China. The purpose of this study was to explore whether the BD Onclarity (Becton, Dickinson and Company)HPV assay differs from the Roche cobas (Roche Molecular Systems)HPV assay, as determined using 944 cervical samples, including 588 with sequencing results. In the nucleic acid assay accuracy verification, the assays showed excellent concordance for detection of HPV16 (κ = 0.93, 95% confidence interval [CI]: 0.89–0.97) and HPV18 (κ = 0.90, 95% CI: 0.83–0.97), and very good concordance for the 12 other high‐risk types (HPV31/33/35/39/45/51/52/56/58/59/66/68, κ = 0.79, 95% CI: 0.75–0.83). The overall agreement for HPV DNA detection between Onclarity and cobas was very good (κ = 0.7755). No difference for ≥CIN2 sensitivity was observed between Onclarity and cobas (both 96.5%), whereas the ≥CIN2 specificity for detection of Onclarity (16.6%, 95% CI: 13.7–19.9) was higher than that of cobas (11.5%, 95% CI: 9.1–14.5). Onclarity exhibited comparable screening performance and triage efficiency compared to cobas in the detection of cervical disease in Chinese women.

The Impact of Specific Sexually Transmitted Pathogens on Cervix: A Prospective Study Based on Cervical Cancer Screening Cohort

ABSTRACT Previous studies showed the association between sexually transmitted infections (STIs) and cervical lesions remains ambiguous. This study was conducted among 8371 women from a screening cohort. Seven specific sexually transmitted pathogens (STPs), including one viral [high‐risk human papillomavirus (hrHPV), low‐risk HPV (lrHPV)], five bacterial [Ureaplasma parvum (UP), Mycoplasma hominis (MH), Ureaplasma urealyticum (UU), Chlamydia trachomatis (CT), and Mycoplasma genitalium (MG)], and one parasitic [Trichomonas vaginalis (TV)] pathogen, were tested by Next Generation Sequencing assay using well‐stored baseline samples. Odds ratios (ORs) for incident cervical lesions with different STPs were calculated by Logistic Regression analysis. Within 3‐year follow‐up, 133 and 72 participants were diagnosed with histopathological cervical intraepithelial neoplasia grade 1 (CIN1) and CIN2+, respectively. The adjusted ORs (aORs) of atypical squamous cells of undetermined significance or worse (ASC‐US+) for women with hrHPV, lrHPV, UP, MH, TV, CT, and MG infections were 2.62 (95% CI: 2.19–3.13), 1.94 (95% CI: 1.55–2.43), 1.48 (95% CI: 1.26–1.74), 1.47 (95% CI: 1.25–1.73), 1.65 (95% CI: 1.27–2.15), 1.26 (95% CI: 0.79–2.01) and 2.33 (95% CI: 1.41–3.85), respectively. The aORs of cytological high‐grade squamous intraepithelial lesions (HSIL) for women with hrHPV, TV, and MG infections were 13.01 (95% CI: 5.78–29.31), 3.48 (95% CI: 1.38–8.75), and 5.87 (95% CI: 1.58–21.77). The aORs of CIN1 for hrHPV, lrHPV, and MH were 6.88(95% CI: 4.79–9.90), 2.04(95% CI: 1.29–3.14), and 1.47(95% CI: 1.02–2.11). The aOR of CIN2+ for women with hrHPV infection was 17.56 (95% CI: 10.31–29.92), no significance was observed for CIN2+ with non‐hrHPV STIs. Specific STP infections were significantly associated with subsequent cervical cytological ASC‐US+ (hrHPV, lrHPV, UP, MH, TV, and MG) and HSIL (hrHPV, TV, and MG). Infection with lrHPV and MH could increase the CIN1 risk in future though no obvious CIN2+ risk elevation was observed.

Performance of a Full-Coverage Cervical Cancer Screening Program Using on an Artificial Intelligence– and Cloud-Based Diagnostic System: Observational Study of an Ultralarge Population

Background The World Health Organization has set a global strategy to eliminate cervical cancer, emphasizing the need for cervical cancer screening coverage to reach 70%. In response, China has developed an action plan to accelerate the elimination of cervical cancer, with Hubei province implementing China’s first provincial full-coverage screening program using an artificial intelligence (AI) and cloud-based diagnostic system. Objective This study aimed to evaluate the performance of AI technology in this full-coverage screening program. The evaluation indicators included accessibility, screening efficiency, diagnostic quality, and program cost. Methods Characteristics of 1,704,461 individuals screened from July 2022 to January 2023 were used to analyze accessibility and AI screening efficiency. A random sample of 220 individuals was used for external diagnostic quality control. The costs of different participating screening institutions were assessed. Results Cervical cancer screening services were extended to all administrative districts, especially in rural areas. Rural women had the highest participation rate at 67.54% (1,147,839/1,699,591). Approximately 1.7 million individuals were screened, achieving a cumulative coverage of 13.45% in about 6 months. Full-coverage programs could be achieved by AI technology in approximately 1 year, which was 87.5 times more efficient than the manual reading of slides. The sample compliance rate was as high as 99.1%, and compliance rates for positive, negative, and pathology biopsy reviews exceeded 96%. The cost of this program was CN ¥49 (the average exchange rate in 2022 is as follows: US $1=CN ¥6.7261) per person, with the primary screening institution and the third-party testing institute receiving CN ¥19 and ¥27, respectively. Conclusions AI-assisted diagnosis has proven to be accessible, efficient, reliable, and low cost, which could support the implementation of full-coverage screening programs, especially in areas with insufficient health resources. AI technology served as a crucial tool for rapidly and effectively increasing screening coverage, which would accelerate the achievement of the World Health Organization’s goals of eliminating cervical cancer.