Search Results (112)

Background: Cytology from cerebrospinal fluid (CSF) is standard-of-care to detect central nervous system (CNS) cancers but suffers from low-sensitivity and lacks associated molecular information, often requiring brain biopsy or resection to obtain. Belay Diagnostics offers analytically and clinically validated CSF-based tests to support the diagnosis and management of primary and secondary CNS cancers. However, the clinical utility (CU) of these assays has not been previously evaluated. Methods: This retrospective study presents a real-world, single institution experience of using the Belay Summit liquid biopsy test for all orders received (n = 123) between October 2024 and September 2025. Clinical information was reviewed for demographics, provisional diagnosis, oncology history, CSF cytology results, and tumor genomic profiling results. The primary endpoint of this study was to evaluate the impact of Belay CSF-based assays on physician diagnosis and treatment decisions. Secondary endpoints included evaluating the clinical performance of the Belay Summit test verses cytology in CNS malignancy detection (sensitivity, specificity, and accuracy). Results: The cohort included 120 patients with suspected or previously diagnosed primary (n = 40) or metastatic (n = 80) CNS tumors; three patients completed longitudinal testing for a total of 123 specimens and test orders. Summit showed higher sensitivity for CNS malignancy (90%) over CSF cytology (12%). The Belay CSF liquid biopsy test demonstrated strong clinical utility and was essential to clinical course pursued in 93% (114/123) of specimens, informing treatment and management decisions. Conclusions: The Belay Summit test provides clinically meaningful information to support physician decision-making for the diagnosis and management of primary and secondary CNS tumors, especially in cases where tissue sampling is infeasible. Full article

Background: Malignant pleural effusion (MPE) represents a frequent and clinically challenging manifestation of advanced malignancy, particularly in metastatic non-small cell lung cancer (NSCLC). Its management requires integration of diagnostic imaging, symptom-directed therapeutic strategies, and, increasingly, molecular profiling technologies. Recent advancements in this field based on liquid medium (so-called liquid biopsy) have achieved a significant increase in sensitivity, enhancing our ability to investigate biofluids and suggesting their potential integration into standard diagnostic practices, far beyond the canonical plasma biopsies. Fluid obtained from MPE after cytological sample centrifugation is rich in cell-free DNA and less susceptible to nucleic acid degradation during processing, improving overall diagnostic accuracy. Methods: This narrative review summarizes current evidence on the clinical management of malignant pleural effusions in patients with metastatic NSCLC, integrating imaging, procedural management, and molecular profiling from a multidisciplinary tumor board perspective. The primary objective was to synthesize contemporary knowledge with particular attention to the feasibility, reliability, and reproducibility of pleural fluid-based molecular testing. Results: MPE poses diagnostic and therapeutic challenges for all members of the multidisciplinary tumor board, traditionally associated with an adverse prognosis. However, recent advances in cytopathology, histopathology, and liquid-based techniques demonstrate that MPE could be an important source of prognostic or predictive information. At the same time, optimal patient management requires careful integration of imaging findings and procedural strategies (such as pleurodesis or indwelling pleural catheters) with individualized systemic therapy selection. Cell-free DNA in pleural effusions is a promising field of exploration and study, potentially suitable for future guideline implementation, after validation in adequately powered studies, contributing to improving patient management, particularly useful in fragile subsets. Conclusions: The management of MPE in advanced NSCLC is evolving toward a multidisciplinary, precision-oriented model that integrates clinical evaluation, imaging, procedural interventions, and molecular testing. Liquid biopsy technology has gained enough analytical robustness and clinical feasibility to be a useful tool in routine analysis. Biofluid-based molecular testing may have outstanding potential, contributing to improving patient management, avoiding repetitive procedures, and optimizing the overall efficiency and cost-effectiveness of diagnostic practices. Moreover, collaborative projects among different specialties help in consolidating trust in the tumor board decision-making process. Full article

Background/Objectives: Co-testing may improve cervical cancer prevention by stratifying women into groups with different absolute risks of CIN2+, CIN3+, and cervical cancer. We evaluated real-world co-testing with cervical cytology and a genotype-specific HPV E6/E7 mRNA assay targeting HPV16, HPV18, and HPV45 (PreTect SEE) in routine clinical practice. Methods: We conducted a retrospective, registry-based cohort study at the Department of Clinical Pathology, University Hospital of North Norway. Eligible co-test samples (liquid-based cytology with concurrent HPV mRNA testing, both with valid results) from routine screening, follow-up, and clinically indicated testing were identified from the laboratory information system and passively followed for worst histological outcome through December 2025. Outcomes were no biopsy/<CIN2, CIN2+, CIN3+, and cervical cancer. Results: Among 116,217 eligible co-test samples (mean age 43.9 years), cumulative risks were 4.4% for CIN2+, 1.5% for CIN3+, and 0.1% for cervical cancer. Baseline HPV mRNA positivity was 3.9%, and cytology was ASC-US+ in 12.2% of samples. Co-testing produced a clear stepwise risk gradient. Double-negative results (NILM/mRNA−; 86.7%) had very low risks (CIN3+ 0.2%; cervical cancer 0.02%). ASC-US+/mRNA− results (9.3%) showed intermediate risks (CIN3+ 4.1%; cervical cancer 0.2%). NILM/mRNA+ results (1.1%) showed substantially higher risks despite normal cytology (CIN3+ 13.0%; cervical cancer 0.5%). Double-positive results (ASC-US+/mRNA+; 2.8%) had the highest risks (CIN3+ 28.5%; cervical cancer 2.3%). Within NILM, mRNA positivity captured 42.7% of CIN3+ cases and 25.0% of cancers. Genotype-specific analyses showed highest risks for HPV16, followed by HPV18 and HPV45. Conclusions: Co-testing with cervical cytology and a 3-type HPV mRNA assay provided strong, clinically interpretable risk stratification and identified a small but high-risk subgroup among women with normal cytology. These findings support genotype-specific HPV mRNA testing as an adjunct to cytology in routine care. Full article

Background: Urothelial carcinoma (UC) is characterized by high recurrence rates and the need for long-term surveillance. Cystoscopy remains the diagnostic gold standard but is invasive, costly, and burdensome for patients. Urine, as a tumor-proximal and non-invasive biospecimen, represents an attractive source for biomarkers enabling screening, diagnosis, risk stratification, and follow-up. Objective: This review summarizes current and emerging urine-based diagnostic approaches for UC, ranging from conventional cytology to advanced molecular technologies, and discusses their clinical utility, limitations, and future perspectives. Methods: A narrative review of the literature was conducted focusing on urine-based diagnostics for UC, including urinary cytology, FDA-approved and investigational protein and DNA/RNA biomarkers, next-generation sequencing (NGS), cell-free DNA (cfDNA), exosomes, and microRNAs. Evidence from clinical validation studies, meta-analyses, and translational research was evaluated. Results: Urinary cytology remains highly specific for high-grade disease but has limited sensitivity for low-grade tumors. Protein- and DNA-based biomarkers have improved sensitivity but often lack sufficient specificity for standalone use. Recent advances in NGS-based assays enable comprehensive detection of tumor-specific genomic alterations in urinary cfDNA, offering high sensitivity for both initial diagnosis and disease monitoring. Exosomes and microRNAs represent promising biomarkers reflecting tumor biology, though standardization and large-scale validation are ongoing challenges. Overall, multimodal approaches combining cytology with molecular assays appear most promising for clinical implementation. Conclusions: Urine-based diagnostics are rapidly evolving toward integrated liquid biopsy platforms capable of transforming UC management. While several assays show strong potential to reduce reliance on cystoscopy, robust prospective validation, cost-effectiveness analyses, and clinical integration strategies are required before widespread adoption. Full article

Automated analysis of liquid-based cervical cytology is increasingly supported by digital microscopy and deep learning. However, model generalization remains challenging due to scanner- and laboratory-induced domain shifts affecting color, texture, and morphology. In this study, we present a robust cell-level classification framework for liquid-based Pap smear cytology based on deep transfer learning, designed to operate under heterogeneous acquisition conditions. We construct a multi-source dataset by integrating three widely used public reference repositories (SIPaKMeD, Herlev, CRIC Cervix) with a proprietary cohort comprising 416 Whole Slide Images (WSIs) collected from two medical centers and digitized using different scanning systems. All labels are harmonized into four Bethesda categories (NILM, ASC-US, LSIL, HSIL), and cell-centered 224 × 224 patches are used as standardized inputs for model development and benchmarking. We evaluate state-of-the-art CNN backbones (ResNet50, EfficientNetB0, VGG16) and perform systematic ablation across data-source combinations to quantify robustness under acquisition variability. Among the evaluated models, ResNet50 yields the best overall performance on the independent test set (accuracy = 0.91; macro-F1 = 0.91), consistently outperforming EfficientNetB0 and VGG16. Importantly, incorporating proprietary multi-center WSI-derived data improves robustness to scanner-induced variation compared to training on public data alone. These findings demonstrate that combining diverse data sources can mitigate domain shift in cell-level cervical cytology classification. While clinically actionable screening requires slide-level aggregation (e.g., MIL-based WSI inference), the proposed classifier provides a robust component that can be integrated into end-to-end WSI screening pipelines in future work. Full article

Background/Objectives: Within human papillomavirus (HPV)-based screening, cytology remains essential for cervical cancer detection while also potentially revealing endometrial pathology. This pilot study aimed to distinguish benign (normal) cases from atypical endometrial hyperplasia (AEH) and endometrial cancer (EC) within atypical glandular cell (AGC) cytology using quantitative analysis of liquid-based cervical cytology. Methods: SurePath and ThinPrep sets included 62 (37 normal, 25 AEH/EC) and 52 (24 normal, 28 AEH/EC) AGC cases, respectively. Semi-automatic QuPath analysis workflow detected cellular clusters; extracted texture, intensity, and geometric features; and produced case-level summaries. A random forest (RF) classifier was used to discriminate AEH/EC from normal cases. Feature subset selection was performed using a beam-search wrapper and joint hyperparameter tuning. Primary performance evaluation comprised stratified 5-fold cross-validation with metrics averaged across these folds. Results: Across both preparations, univariable analyses showed moderate discrimination overall which improved post-menopause. For SurePath and ThinPrep, the highest 10 areas under the curve (AUCs) were 0.701–0.773 (improving to 0.798–0.841 post-menopause) and 0.740–0.778 (improving to 0.832–0.884 post-menopause), respectively. Machine-learning RF models improved performance beyond univariable baselines. Cross-validated AUCs for SurePath and ThinPrep were 0.805 (95% confidence interval [CI], 0.683–0.927) and 0.887 (95% CI, 0.787–0.987), respectively. Features associated with higher AUCs differed between SurePath and ThinPrep, indicating platform-specific signals. Conclusions: Quantitative analysis of routine cervical cytology can augment expert reviews to help distinguish endometrial lesions among AGCs, particularly post-menopause. These software-based readouts can fit within existing workflows and may improve triage when morphology is subtle, including scenarios with HPV-negative screening results. Full article

Background: Human papillomavirus (HPV)-based screening has substantially improved sensitivity for cervical cancer detection but remains limited by low specificity, leading to unnecessary colposcopy referrals. MicroRNAs (miRNAs) represent promising biomarkers for improving triage of HPV-positive women. This study evaluated the diagnostic and regulatory roles of selected miRNAs in cervical lesion progression using liquid-based cytology (LBC) specimens. Methods: Expression of six biologically relevant miRNAs (miR-15a-5p, miR-16-5p, miR-20b-5p, miR-155-5p, miR-34a-5p, and miR-140-3p) was analyzed across NILM, CIN II, CIN III, and cervical cancer (CC) samples. All miRNA analyses were performed using residual cellular material derived from the same liquid-based cytology (LBC) specimens collected during the HPV screening visit, without requiring any additional sampling prior to colposcopy. Diagnostic performance was assessed using ROC analysis. To capture regulatory dynamics beyond expression magnitude, correlation, and differential correlation (Δρ), network analyses were applied. Results: Stage-dependent changes in miRNA expression were observed across disease categories; however, expression magnitude alone did not fully explain diagnostic performance. Upregulated miRNAs, particularly miR-16-5p, miR-20b-5p, and miR-155-5p, demonstrated high diagnostic accuracy for distinguishing NILM from high-grade lesions and invasive cancer. In contrast, downregulated miRNAs showed limited diagnostic utility. Correlation analyses revealed progressive remodeling of miRNA co-expression networks, with the most pronounced changes occurring during the CIN II–to–CIN III transition. Notably, miRNAs with strong diagnostic performance did not uniformly function as network hubs, indicating distinct roles as biomarkers versus regulators of network dynamics. Conclusions: Cervical lesion progression is characterized not only by changes in miRNA expression levels but also by stage-specific reorganization of miRNA regulatory networks. Integrating diagnostic performance with network-level analysis enables improved identification of clinically robust triage markers and provides additional insight into regulatory instability associated with progression. Full article

HPV DNA–positive women with ASC-US/LSIL cytology represent a heterogeneous risk group in cervical screening and require efficient triage. We evaluated a genotype-specific 7-type HPV E6/E7 mRNA assay (PreTect HPV-Proofer 7; types 16/18/31/33/45/52/58) in a real-world quality-assurance cohort at Nordland Hospital (Bodø, Norway). Among HPV-positive women with ASC-US/LSIL reflex cytology, 225 had sufficient residual liquid-based cytology material and a valid mRNA result; 175 had complete follow-up (2022–2025) and were included. Overall, 44.6% (78/175) were mRNA-positive (ASC-US 45.2%; LSIL 43.3%). For CIN2+, sensitivity was 63.4%, specificity 61.2%, PPV 33.3%, and NPV 84.5%; CIN2+ risk was 33.3% in mRNA-positive versus 15.5% in mRNA-negative women (RR 2.16, 95% CI 1.23–3.78). For CIN3+, risk was 14.1% versus 6.2%. Genotype-specific PPVs were highest for HPV33, HPV18, HPV16, and HPV31. In a referral simulation, mRNA-guided triage reduced baseline colposcopy referrals by 55% and decreased colposcopies per detected CIN2+ by ~30%, while 15 CIN2+ and 6 CIN3+ occurred in the mRNA-negative group and would be expected to be detected at 12-month follow-up among women with persistent HPV positivity. Genotype-aware HPV E6/E7 mRNA triage improves risk stratification and may increase screening efficiency. Full article

Background: Vaccination against human papillomaviruses (HPVs) and cervical cancer screening represent effective tools for preventing this neoplasia, but access to health services is limited for vulnerable women. We investigated prevalence of high-risk HPV and abnormal cervical cytology, as well as knowledge about HPV and the HPV vaccine, among homeless and migrant women in Rome, Italy. Methods: Cytologic samples in PreservCyt (Hologic) were employed for liquid-based cytology (ThinPrep Processor 5000, Hologic) and high-risk HPV DNA testing (Xpert HPV assay, Cepheid). Socio-demographic data, anamnestic, and behavioral data were retrieved from electronic archives. A questionnaire was employed to assess knowledge about HPV and HPV vaccination. Results: A total of 134 women were included (median age: 43 years; interquartile range, IQR: 34–50), mostly coming from Central–South America (69, 51.5%). Of the 127 cytologic specimens collected, one (0.8%) was invalid for the HPV test and five (3.9%) were unsatisfactory for the morphological evaluation. High-risk HPV positivity was found in 18 women of the 126 women with a valid HPV test (14.3%). A total of 10 women of the 122 women with an adequate cytology (8.2%) had abnormal cytology. Overall, 57/134 women (42.6%) had never heard of HPV or were unsure about it. Only 29 of the 77 women who had heard of HPV (37.7%) knew of the HPV vaccine, and only 2 had been vaccinated in the entire study group (1.5%). Conclusions: Tailored preventive strategies and comprehensive information campaigns should be developed and implemented to enhance awareness of HPV infection and actively promote vaccination among women in vulnerable groups. Full article

Cervical cancer remains a significant global health burden, disproportionately affecting women in low- and middle-income countries despite being preventable. Since 2018, rapid advances in molecular profiling, immunotherapy, refinement of minimally invasive surgery, and targeted therapeutics have transformed diagnostic and therapeutic paradigms. This narrative review synthesizes clinical and translational progress across the continuum of care from 2018 to 2025. We summarize the evolving landscape of precision screening—including HPV genotyping, DNA methylation assays, liquid biopsy, and AI-assisted cytology—and discuss their implications for global elimination goals. Surgical management has shifted toward evidence-based de-escalation with data from SHAPE, ConCerv, and ongoing RACC informing fertility preservation and minimally invasive approaches. For locally advanced disease, KEYNOTE-A18 establishes pembrolizumab plus chemoradiation as a new curative standard, while INTERLACE underscores the benefit of induction chemotherapy. In the metastatic setting, survival outcomes have improved with the integration of checkpoint inhibitors (KEYNOTE-826, BEATcc, EMPOWER-Cervical 1), vascular-targeted therapies, and antibody–drug conjugates, including tisotumab vedotin and emerging HER2 and TROP-2–directed agents. We further highlight emerging biomarkers—PD-L1, TMB, MSI status, HPV integration patterns, APOBEC signatures, methylation classifiers, ctHPV-DNA—and their evolving role in treatment selection and surveillance. Future directions include neoadjuvant checkpoint inhibition, PARP-IO combinations, HER3-directed ADCs, DDR-targeted radiosensitizers, HPV-specific cellular therapies, and AI-integrated precision medicine. Collectively, these advances are reshaping cervical cancer care toward biologically individualized, globally implementable strategies capable of accelerating WHO elimination targets. Full article

Cervical cancer (CC) can be prevented through continuous screening and the timely detection of cervical intraepithelial neoplasia (CIN) using immunohistochemistry techniques to identify biomarker expressions. In a previous study, we proposed nuclear REST loss as a biomarker in precancerous lesions and CC; however, no validated antibodies are available for detecting REST in cytology or cervical tissues. Although we have developed an IgM-type anti-REST monoclonal antibody capable of detecting REST in liquid-based cytology cells, it was not useful for the detection of REST in cervical tissues by immunohistochemistry. The main objective of this study is to generate single-chain variable fragments (scFvs) for the clinical evaluation of REST in cervical tissues from women with CIN and CC. Using RNA from an IgM-producing hybridoma anti-REST, we conducted RT-PCR and PCR to obtain the coding sequences for the variable regions of the heavy and light chains. These sequences were joined with a linker to create a single-chain antibody. The scFv was then cloned into the pSyn1 vector, expressed in E. coli TG1, and purified through chromatography. Subsequently, it was characterized using immunological methods to assess its biological activity and employed to evaluate REST expression in cytological samples and cervical tissues. The anti-REST scFv represents an innovative detection tool that retains the antigen recognition of the parental IgM while overcoming its size limitation, enabling tissue penetration and detection of REST in cervical samples. Its application facilitates the identification of REST in cervical samples, reinforcing REST’s potential as a diagnostic biomarker for CC and CIN. Full article

In Norway, organized cervical cancer screening was cytology-based until 2023, and women screened in 2013–2015 were largely unvaccinated. We conducted a retrospective, population-based cohort study to assess whether co-testing with a 3-type HPV mRNA assay improves detection of high-grade cervical lesions in women < 40 years. Among 11,395 women screened in Northern Norway and followed for 8–10 years, 2807 formed a co-testing cohort (ThinPrep cytology plus PreTect SEE; HPV16/18/45) and 8588 formed a cytology-only cohort. The endpoint was histologically confirmed CIN2+. Sensitivity for CIN2+ was 63.7% with cytology alone and 71.0% with co-testing (absolute +7.3 percentage points; p = 0.034). In the co-testing cohort, HPV mRNA was detected in 10.2% of women, of whom 46.0% developed CIN2+, while CIN2+ risk in HPV mRNA-negative women was 5.2%. Co-testing produced wide risk gradients: CIN2+ risk was 58.3% in double-positive women (HPV mRNA-positive and ASC-US+) and 3.3% in double-negative women (HPV mRNA-negative and normal cytology), with no cervical cancers observed in the latter group. In this cytology-based, largely unvaccinated setting, co-testing with a 3-type HPV mRNA assay improved detection performance and long-term risk stratification in women < 40 years, supporting its use as a quality-assurance and triage tool within organized screening programs. Full article

Background/Objectives: Atypical cytological findings in cervical screening, such as ASC-US, ASC-H, and AGC, present a clinical challenge due to their variable risk of underlying high-grade lesions. The precise stratification of this risk is crucial for effective patient management. This study aimed to correlate Bethesda cytology categories with HPV genotyping, including viral load, and histological follow-up to improve risk prediction for cervical intraepithelial neoplasia grade 2 or worse (CIN2+). Materials and Methods: In this retrospective single-center study, we analyzed 407 patients with cytological reports of ASC-US, ASC-H, or AGC. All patients underwent HPV DNA testing with genotyping for 21 types, with viral load quantification for HPV16/18, and subsequent histological verification. Statistical analyses included non-parametric tests, correlation analysis, and multivariate logistic regression to identify independent predictors of CIN2+. Results: The prevalence of CIN2+ differed significantly among the cytological categories: 23.2% in ASC-US, 47.3% in ASC-H, and 19.5% in AGC. ASC-H and a high HPV16 viral load were identified as independent predictors of CIN2+ in the multivariate analysis. An ASC-H result increased the probability of CIN2+ by 2.5 times (aOR = 2.51; 95% CI: 1.28–4.94). For each 1 log10 increase in HPV16 viral load, the risk of CIN2+ increased by 30% (aOR = 1.30; 95% CI: 1.16–1.46). Stratification of ASC-US cases by HPV16 status revealed a dramatically higher positive predictive value (PPV) for CIN2+ in HPV16-positive patients (66%) compared to HPV16-negative patients (12.6%). The AGC category showed the strongest association with glandular pathology, including adenocarcinoma in situ. Conclusions: The combination of cytological findings and HPV16 viral load provides a powerful model for risk stratification. An ASC-H result is a strong independent risk marker, while the clinical significance of ASC-US is fundamentally determined by HPV16 status. These findings advocate for a risk-based management algorithm that integrates liquid-based cytology with extended HPV genotyping and viral load assessments to optimize patient triage and follow-up. Full article

Colorectal cancer (CRC) remains a leading cause of cancer-related mortality, and detecting circulating tumor cells (CTCs) is crucial for early diagnosis and metastasis monitoring. Conventional staining-based cytology is costly, time-consuming, and often compromises sample integrity. In this study, we employed a combined digital holography (DH) and fluorescence imaging approach to develop a virtual staining framework for transforming quantitative phase imaging (QPI) data into interpretable pseudo-stained images. To the best of our knowledge, this is the first application of such a framework to colorectal cancer CTC detection. In our experiments, green fluorescent protein (GFP)-labeled HCT116 cells—generated through lentiviral transfection—were mixed with peripheral blood mononuclear cells (PBMCs) to create training datasets. The trained network achieved 99% classification accuracy and demonstrated strong generalization to unseen donors. This DH–fluorescence-based virtual staining method preserves cell integrity while enabling rapid, label-free, and low-cost liquid cytology diagnostics, highlighting its potential for non-invasive cancer detection and monitoring. Full article

Background/Objectives: Cervical cancer is one of the most frequent malignancies among women in Kazakhstan, where human papillomavirus (HPV) vaccination was initiated in 2024. Despite the implementation of vaccination and cytology-based screening programs, diagnostic limitations remain, and local evidence linking HPV infection to clinical outcomes is scarce. This study aimed to evaluate the correlation between HPV status, cervical cytology results, colposcopic impression, and biopsy results in a non-vaccinated female population. Methods: A cross-sectional study was conducted at the University Medical Center, Astana, between November 2024 and March 2025. A total of 396 women of reproductive age were enrolled. Cervical samples underwent liquid-based cytology and high-risk HPV testing with the RealBest assay. Colposcopy was performed following abnormal cervical cytology results, and colposcopy-guided biopsies were obtained where indicated. Sociodemographic characteristics were assessed, and associations between HPV genotype and clinical outcomes were analyzed using descriptive and inferential statistics. Results: HPV infection was detected in 140 women (35.4%). HPV-16 was the most common genotype (11.4%), followed by HPV-52 (6.6%) and HPV-33 (5.3%). Among 198 women evaluated by colposcopy, abnormal findings were observed in 72.2%, with HPV-16 showing a significant association with higher-grade abnormalities (p < 0.001). Biopsies were available for 40 participants: 12 had CIN I, 12 had CIN II, 10 had CIN III, and 4 had carcinoma in situ. HPV-16 was the only genotype significantly linked to CIN II/III lesions. Conclusions: HPV-16 was strongly associated with abnormal colposcopic findings and high-grade histology, underscoring its oncogenic importance. The prevalence of HPV-52 and HPV-33 further supports the need for HPV nonavalent vaccination. These findings highlight the importance of HPV-based screening, genotype-specific triage, and expanded vaccination to reduce cervical cancer incidence in Kazakhstan. Full article