ABSTRACT Introduction Cervical precancerous lesions and cancer are caused by persistent oncogenic high-risk human papillomavirus (HR-HPV). Molecular HPV DNA testing has recently replaced cytology as the preferred primary screening method because of its superior sensitivity and negative predictive value. Nevertheless, its limited specificity can result in unnecessary follow-up procedures. Therefore, the identification of surrogate biomarkers capable of distinguishing transient infections from persistent and clinically relevant ones is essential for improving triage and risk stratification. To summarize the available evidence, a comprehensive literature search was performed across PubMed/MEDLINE, Embase, Scopus, and Web of Science databases up to December 2025, identifying studies that evaluated HR-HPV viral load in relation to cervical intraepithelial lesions and cancer progression. Areas Covered Oncogenic HPV viral load, defined as the quantity of HPV genomes in a cervical sample or biopsy tissue, may constitute an attractive biomarker for the progression of HPV-related cervical lesions. Indeed, HPV viral load levels are positively correlated with the persistence of cervical HR-HPV, leading to an increased risk of high-grade lesions and invasive cervical cancer. Furthermore, cervical HPV viral load quantification provides useful prognostic information regarding disease severity, therapeutic response, and recurrence following treatment of cervical lesions. Expert Opinion The recent standardization and validation of available multiplex real-time PCR techniques for HPV detection and quantification supports the clinical integration of HPV viral load as a surrogate biomarker. Incorporating viral load assessment into cervical screening and management algorithms could significantly enhance diagnostic precision, enable more personalized follow-up, and guide therapeutic decisions for women affected by HR-HPV-associated precancerous and cancerous cervical diseases.

ABSTRACT Introduction Helicobacter pylori (H. pylori) infection is widespread globally, affecting more than half of the world’s population. However, traditional detection methods, which have been used for decades, have several limitations, such as invasiveness and the requirement for endoscopy to acquire biopsy samples. Given the severity and complexity of the diseases caused by H. pylori, biosensors provide a practical solution for H. pylori detection, offering a valuable approach to simple, accurate, rapid, and highly specific identification of stomach cancer in its early stages. Areas covered This review paper provides a comprehensive overview of various biosensors categorized into distinct sections based on their classification. Different electrochemical sensors, aptamer-based, optical, whole-cell, enzyme-based, and immunosensors that have already been developed for the detection of various biomarkers of H. pylori are summarized. Expert opinion Future research should focus on developing more reliable, easier, cost-effective, and valid H. pylori biomarker-based biosensors to enhance diagnostics and therapeutic outcomes. There is also a need to commercialize these biosensors, enabling individuals to detect disease themselves at home or facilitating real-time detection. The successful translation of these biosensors from laboratory settings to commercial use necessitates a series of defined steps, which warrant immediate prioritization.

ABSTRACT Introduction Since 1993, when a polymorphic CAG trinucleotide repeat was established as the cause of Huntington’s disease (HD), fragment analysis has been the first-tier test in genetic diagnosis of HD. However, in recent years it has become increasingly clear that, in addition to the repeat length itself, sequence variations within the HTT repeat and elsewhere in the genome also play a crucial role in the onset and clinical presentation of the disease. Areas covered Sequence interruptions or non-canonical repeat motifs have historically been difficult to identify, as PCR cannot distinguish an interrupted repeat tract when assessed by capillary electrophoresis. However, these non-canonical interruptions may directly affect the phenotype or result in an underestimation of the uninterrupted CAG repeat length. While next-generation sequencing (NGS) with short read lengths has revolutionized genetic diagnostics, it faces limitations in diagnosing repeat expansion diseases. The literature search methodology involved a review of relevant studies and clinical data. Expert opinion In contrast, long-read sequencing (LRS) generates read lengths of over 10 kb and can thus overcome the limitations associated with short reads. This enables detailed investigation of repeat sequences and the identification of complex patterns of alternating sequence motifs, potentially providing additional clinically relevant information.

ABSTRACT Introduction Ovarian cancer is a leading cause of cancer-related death in women, primarily due to nonspecific symptoms and diagnosis at an advanced stage. Improving outcomes through earlier diagnosis and effective population screening remains a global priority. Context Existing modalities ,including CA125 and transvaginal ultrasound (TVUS), lack sufficient specificity and sensitivity, and no screening strategy has demonstrated a reduction in mortality. Multivariate Index Assays (MIAs), that combine multiple biomarker results into a single risk score, have emerged as a promising approach for differential diagnosis of adnexal masses, early detection, and screening. Areas covered This review summarizes current literature and clinical trial data on MIAs in ovarian cancer, describing their advantages over single biomarkers, their potential to improve patient outcomes, and their role in personalized medicin. Successful clinical adoption of MIAs will depend on well-designed prospective studies that robustly validate their clinical utility. Expert opinion/commentary It is also critical to clearly distinguish between diagnostic and screening applications, as each requires different performance characteristics, regulatory pathways, and target populations, and no ovarian cancer screening test is currently approved for routine use.

ABSTRACT Introduction MUC4 is overexpressed in gastric cancer and its precursor lesions and plays an essential role in disease progression. Area covered In this review, we describe the diagnostic and prognostic value of MUC4 expression in gastric cancer. First, we provide a summary of gastric cancer pathogenesis and its intricate relationship with mucin expression. We will then provide an extensive description of MUC4 expression in gastric cancer and its diagnostic and therapeutic significance. Expert opinion Apart from its protective role in normal physiology, MUC4 contributes to disease progression and metastasis in several malignancies, including gastric cancer. MUC4 is expressed at negligible levels in normal gastric mucosa. However, some patients with intestinal metaplasia and gastric cancer have high levels of MUC4 expression, particularly in the pit cell compartment. The expression of MUC4 in metaplastic pit cells is associated with increased proliferation, inflammation, and epithelial-to-mesenchymal transition. Hence, evaluating MUC4 expression in precursor lesions may help stratify disease and identify cases in which curative surgery should be performed. MUC4 expression in gastric cancer is associated with disease aggressiveness, lymphatic invasion, and nodal metastasis. Due to its role in disease pathobiology, it is a promising target for the diagnosis and therapy of gastric cancer.

ABSTRACT Introduction Cholangiocarcinoma (CCA) is an uncommon yet aggressive malignancy often diagnosed at advanced stages. Its management is challenged by significant molecular heterogeneity and limited treatment options. Advances in next-generation sequencing (NGS) have identified actionable alterations, such as FGFR2 fusions, thereby facilitating a precision oncology approach for CCA management. Areas Covered This review consolidates current evidence and expert insights on molecular profiling in CCA. It examines the histopathological subtypes and addresses diagnostic challenges associated with their diagnosis. Critical pre-analytical factors, including biopsy techniques, tissue handling, and tumor heterogeneity, are discussed in relation to their impact on molecular testing. The review also evaluates DNA-based versus RNA-based NGS methodologies, highlighting their strengths and limitations in detecting complex genomic alterations. The role of liquid biopsy as a minimally invasive tool for dynamic tumor monitoring is also explored. Expert Opinion The routine integration of molecular profiling for CCA requires the best histopathological diagnosis and pre-analytical preparation practices. Diagnostic workflows should prioritize meticulous tissue handling to ensure robust molecular analyses to avoid tissue exhaustion and preserve the integrity of nucleic acids. Employing DNA plus RNA sequencing platforms, supported by molecular tumor boards, is recommended to enhance patient stratification and guide therapeutic decision-making in CCA.

ABSTRACT Introduction Pathogenesis of large B-cell lymphomas (LBCL) and follicular lymphomas (FL) is a multistep process associated with the development of diverse DNA alterations and consequent deregulation of critical cellular processes. Detection of tumor-associated mutations within non-tumor compartments (mainly plasma) is the basis of the ‘liquid biopsy’ concept. Apart from tumor mutational profiling, quantitative analysis of circulating tumor DNA (ctDNA) allows longitudinal assessment of tumor burden. ctDNA-based technologies provide a new tool for tumor diagnostics and treatment personalization. Areas covered Our review provides a comprehensive overview and summary of available ctDNA studies in LBCL and FL. The accuracy of ctDNA-based detection of lymphoma-associated DNA alterations is correlated to known LBCL and FL molecular landscape. Additionally, we summarized available evidence that supports and justifies the clinical use of ctDNA for lymphoma risk stratification, treatment response evaluation, and treatment response-adapted therapy. Lastly, we discuss other clinically important ctDNA applications: monitoring of lymphoma clonal evolution within resistance and/or relapse development and utilization of ctDNA for diagnostics in non-blood fluids and compartments (e.g. cerebrospinal fluid in primary CNS lymphomas). Expert opinion Despite certain challenges, including methodological standardization, ctDNA holds promise to soon become an integral part of lymphoma diagnostics and treatment management.

ABSTRACT Introduction Saliva has emerged as an important biological fluid for diagnostics, particularly hormone analysis. Its noninvasive collection and accessibility make it a compelling alternative to traditional blood-based diagnostics, enabling detection of biomarkers reflecting physiological and pathological conditions. Areas covered This review examines hormones measurable in saliva, including cortisol, testosterone, progesterone, estradiol, dehydroepiandrosterone (DHEA), and others. It highlights methods such as Enzyme-Linked Immunosorbent Assay (ELISA), Radioimmunoassay, and Liquid Chromatography coupled with Tandem Mass Spectrometry (LC-MS/MS) for hormonal analysis, focusing on their sensitivity and challenges. The discussion addresses the advantages and limitations of saliva as a diagnostic medium, including practicality and susceptibility to external influences. Clinical applications are explored, including stress monitoring, hormonal dysfunction diagnosis, and applications in personalized medicine. Expert opinion Salivary diagnostics holds significant potential in clinical and research contexts, particularly for hormone analysis. Despite challenges such as hormonal variability and technical limitations, advances are steadily overcoming these barriers. The noninvasive and accessible nature of saliva collection positions it as a promising medium for diagnostic innovation. Continued research, coupled with standardization of techniques, will be critical to fully harnessing saliva-based diagnostics for advancing personalized medicine, influencing the detection, diagnosis, and prognosis of certain conditions.

ABSTRACT Background Ameloblastic carcinoma (AC), malignancy originating from the odontogenic epithelium, shows histological overlap with ameloblastoma (AM). In order to unravel mechanisms driving AC, it is imperative to understand the molecular distinction between these two entities. This systematic review aims to highlight molecular and immunohistochemical markers involved in the pathogenesis of AC and to distinguish it from its histological mimic, AM. Research design and methods Literature search across three databases including PubMed, Web of Sciences, and Scopus was carried out from year 1999 to 2023 for original human case control studies involving AM, AC, and controls as study groups. Various biological markers studied in the literature were grouped based on principal molecular pathways. Joanna Briggs Institute (JBI), a critical appraisal tool for case control studies, was used to assess the risk of bias (RoB). Results Out of the 277 studies identified during the initial search, 28 studies were found eligible. These studies reported expression of various immunohistochemical (IHC), genetic and epigenetic markers in AC, AM, and controls through immunohistochemistry and gene sequencing. Conclusion Stem cells, epigenetics, and growth factors define the pathways involved in pathogenesis of AC and may prove to be a potential therapeutic target in the future.