The Journal of Liquid Biopsy最新文献

{"title":"Novel applications of liquid Biopsy: Comprehensive methodology for circulating biomarker exploration in peripheral blood","authors":"Caterina De Rosa ,&nbsp;Luisa Amato ,&nbsp;Annalisa Ariano ,&nbsp;Sara Capaldo ,&nbsp;Daniela Esposito ,&nbsp;Hamid Heydari Sheikhhossein ,&nbsp;Alessia Salzillo ,&nbsp;Alessandra Di Liello ,&nbsp;Concetta Tuccillo ,&nbsp;Martina Cesarano ,&nbsp;Daniela Frezzetti ,&nbsp;Rosa Carmelingo ,&nbsp;Antonella De Luca ,&nbsp;Antonio Gambardella ,&nbsp;Virginia Tirino ,&nbsp;Fortunato Ciardiello ,&nbsp;Floriana Morgillo ,&nbsp;Federica Papaccio ,&nbsp;Alberto Servetto ,&nbsp;Viviana De Rosa ,&nbsp;Carminia Maria Della Corte","doi":"10.1016/j.jlb.2025.100307","DOIUrl":"10.1016/j.jlb.2025.100307","url":null,"abstract":"<div><div>The liquid biopsy (LB) represents a minimally invasive method for cancer screening that has been introduced in clinical practice for over a decade and that can accelerate treatment response assessment. LB allows the analysis of tumor cells or tumor-derived products (e.g. cell-free circulating nucleic acids, extracellular vesicles, and proteins) released from primary or metastatic tumor lesions into blood or other body fluids. In the era of immune-oncology, recent evidence indicates that tumor-specific immune responses can be detected in peripheral immune cells. The improvement of knowledge and the standardization of the isolation methods of these techniques will allow the detection and characterization of circulating tumor and immune biomarkers at an early stage as innovative tools to predict response to therapies. Nowadays, the analysis of peripheral blood mononuclear cells (PBMCs), circulating tumor cells (CTCs), peripheral blood-derived extracellular vesicles (EVs) and circulating tumor RNA (ctRNA) remains under-developed even if these non-invasive techniques can provide the complete genetic landscape of tumors and allow systematic tracking of cancer evolution. In addition, the evaluation of blood circulating cytokines, and early dynamics changes in the PBMCs of patients with solid tumors represent a promising area of research. Here, we present a comprehensive methodological framework for the evaluation of innovative peripheral blood-derived biomarkers. We also address the current challenges in isolation methods and analysis of PBMC, CTC, EVs and TEPs which are crucial for structuring the large amount of comprehensive information obtained from such samples, with the aim of advancing the translational cancer field.</div></div>","PeriodicalId":101235,"journal":{"name":"The Journal of Liquid Biopsy","volume":"9 ","pages":"Article 100307"},"PeriodicalIF":0.0,"publicationDate":"2025-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144562984","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Blood and urine-based biomarkers in prostate cancer: Current advances, clinical applications, and future directions","authors":"Felice Crocetto ,&nbsp;Michele Musone ,&nbsp;Stefano Chianese ,&nbsp;Paolo Conforti ,&nbsp;Gaetano Digitale Selvaggio ,&nbsp;Vincenzo Francesco Caputo ,&nbsp;Roberto Falabella ,&nbsp;Francesco Del Giudice ,&nbsp;Carlo Giulioni ,&nbsp;Angelo Cafarelli ,&nbsp;Giuseppe Lucarelli ,&nbsp;Gian Maria Busetto ,&nbsp;Matteo Ferro ,&nbsp;Biagio Barone ,&nbsp;Fabio Zattoni ,&nbsp;Daniela Terracciano","doi":"10.1016/j.jlb.2025.100305","DOIUrl":"10.1016/j.jlb.2025.100305","url":null,"abstract":"<div><div>Prostate cancer (PCa) is one of the most prevalent malignancies in men, characterized by high clinical and molecular heterogeneity. Despite the widespread use of prostate-specific antigen (PSA) for diagnosis and monitoring, its limited specificity and sensitivity necessitate the development of more accurate biomarkers. This review provides a comprehensive overview of current and emerging diagnostic, prognostic, and predictive biomarkers in PCa, highlighting their clinical applications and future perspectives. PSA, though historically central in PCa screening, lacks tumor specificity, often leading to unnecessary biopsies or missed aggressive cancers. Recent blood-based biomarkers, such as the Prostate Health Index (PHI) and 4Kscore, improve specificity by integrating PSA isoforms or kallikrein protein levels with clinical parameters. Urine-based biomarkers like PCA3 and SelectMDx further enhance diagnostic precision, particularly in distinguishing high-grade tumors, and show potential in active surveillance settings. Prognostic markers such as Bcl-2, Ki-67, and EZH2, alongside genetic alterations like MCM7 and 8q gain, help stratify patients by tumor aggressiveness and risk of recurrence. Liquid biopsies, including circulating tumor DNA (ctDNA) and circulating tumor cells (CTCs), offer non-invasive alternatives for molecular profiling, especially in metastatic castration-resistant PCa (mCRPC), and can identify actionable alterations such as BRCA1/2 or ATM mutations. Emerging technologies such as machine learning and single-cell omics are reshaping biomarker discovery. Artificial intelligence-driven models, like the replication stress signature (RSS), show promise in predicting relapse and therapeutic response. Single-cell RNA sequencing and spatial transcriptomics have deepened our understanding of PCa heterogeneity, tumour microenvironment, and resistance mechanisms. Furthermore, novel biomarkers, including exosome RNAs and immune-related markers (PD-L1, SOX2, TcellinfGEP), offer insights into tumour progression and immunotherapeutic potential. The urinary and gut microbiomes are also being explored for their diagnostic and prognostic roles in PCa. In conclusion, integrating advanced molecular tools and biomarker-guided platforms into clinical practice can significantly enhance early detection, personalized treatment, and monitoring in prostate cancer, paving the way for precision oncology.</div></div>","PeriodicalId":101235,"journal":{"name":"The Journal of Liquid Biopsy","volume":"9 ","pages":"Article 100305"},"PeriodicalIF":0.0,"publicationDate":"2025-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144321598","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Liquid biopsy as a potential tool for diagnosis and clinical monitoring of cutaneous and uveal melanoma","authors":"Letícia Silva Ferraz ,&nbsp;Alana Silveira Oliveira Souza ,&nbsp;Tiago Rodrigues ,&nbsp;Mauro Jorge Cabral-Castro","doi":"10.1016/j.jlb.2025.100306","DOIUrl":"10.1016/j.jlb.2025.100306","url":null,"abstract":"<div><div>Melanoma is the most aggressive type of skin cancer, characterized by high metastatic potential and high mortality rates. Currently, diagnosis and mutational status assessment involves the excisional tissue biopsy followed by anatomopathological and immunohistochemical analyses. However, this technique is invasive and does not assess the molecular heterogeneity of the tumor. Also, it cannot be applied for the continuous monitoring of the disease. Gathering this information is essential for tumor staging, as well as for helping to select the most appropriate therapeutic approach for the patient. To overcome these limitations, liquid biopsy has arisen as an innovative diagnostic tool, which is a molecular determination technique for assessing tumor heterogeneity. Liquid biopsy is defined as the sampling of circulating biomarkers from tumor cells in various biological matrices, including circulating tumor cells (CTCs), circulating tumor DNA (ctDNA), circulating tumor RNA, extracellular vesicles (EVs), and proteins. In this review, we will discuss the potential of liquid biopsy for the clinical assessment of cutaneous and uveal melanoma, evaluating advantages and some parameters, such as efficiency and specificity. To this end, a bibliographical survey was carried out in the main search databases for scientific articles in the health area, published between March 2019 and October 2024, using the keywords “melanoma”, “biomarker”, and “liquid biopsy”, and 56 articles were found. All these articles highlight the potential of liquid biopsy for personalized treatment of melanoma, early screening, diagnosis and prognosis, detection of relapses, monitoring of metastases, and selection of appropriated treatments. However, additional randomized controlled clinical trials comparing the use of liquid biopsy with the diagnostic gold standard are still needed to validate and evaluate the benefits of its use in clinical practice.</div></div>","PeriodicalId":101235,"journal":{"name":"The Journal of Liquid Biopsy","volume":"9 ","pages":"Article 100306"},"PeriodicalIF":0.0,"publicationDate":"2025-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144281196","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"International society of liquid biopsy (ISLB) perspective on minimal requirements for ctDNA testing in solid tumors","authors":"Nicola Fusco ,&nbsp;Konstantinos Venetis ,&nbsp;Francesco Pepe ,&nbsp;Omshree Shetty ,&nbsp;Silvia Calabuig Farinas ,&nbsp;Simon Heeke ,&nbsp;Julia V. Burnier ,&nbsp;Simon J. Patton ,&nbsp;Ellen Heitzer ,&nbsp;Giovanni Nigita ,&nbsp;Paul Hofman ,&nbsp;Maria Jose Serrano ,&nbsp;Massimo Cristofanilli ,&nbsp;Eloisa Jantus-Lewintre ,&nbsp;David R. Gandara ,&nbsp;Christian Rolfo ,&nbsp;Umberto Malapelle","doi":"10.1016/j.jlb.2025.100301","DOIUrl":"10.1016/j.jlb.2025.100301","url":null,"abstract":"<div><div>Circulating tumor DNA (ctDNA) testing has transformed precision oncology by enabling the non-invasive detection of actionable mutations. To facilitate broader clinical adoption and improve testing accuracy, standardized quality criteria must be clearly defined and universally implemented. The International Society of Liquid Biopsy (ISLB) established the Quality Control and Accreditation Committee to develop consensus-based minimal standards for ctDNA analysis in oncology. Ensuring reliable and reproducible ctDNA testing necessitates standardization across the pre-analytical, analytical, and post-analytical phases. Key considerations include appropriate blood collection, efficient cfDNA isolation and purification, thorough assay validation, and precise data interpretation. The ISLB is committed to leading collaborative efforts among laboratories, regulatory bodies, and professional organizations to advance standardization and ensure high-quality ctDNA testing worldwide. Through initiatives led by the Quality Control and Accreditation Committee, educational programs, and multidisciplinary stakeholder workshops, ISLB provides a structured framework to promote standardization and foster innovation. By addressing current challenges and advocating for robust quality standards, ctDNA testing can reach its full potential in advancing personalized cancer care, enabling more precise and timely interventions for patients. This manuscript provides the first global initiative for quality control in liquid biopsy, presenting the ISLB perspective on minimal requirements for ctDNA testing in solid tumors.</div></div>","PeriodicalId":101235,"journal":{"name":"The Journal of Liquid Biopsy","volume":"8 ","pages":"Article 100301"},"PeriodicalIF":0.0,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144177796","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Utility of combined solid and liquid biopsy for molecular profiling in lung adenocarcinoma: Insights from a real-world case","authors":"Bharat Bhosale ,&nbsp;Gunj Bafna ,&nbsp;Paridhy Vanniya Subramanyam ,&nbsp;Kapil Salgia ,&nbsp;Sadia Afreen ,&nbsp;Jinumary John ,&nbsp;Vyomesh Javle ,&nbsp;Kritika Verma ,&nbsp;N.R. Naavarasi ,&nbsp;Sreekanth Reddy Peddagangannagari ,&nbsp;Akhil Gorla ,&nbsp;Giridharan Periyasamy ,&nbsp;Kshitij Rishi ,&nbsp;Hitesh Goswami ,&nbsp;Vidya H. Veldore","doi":"10.1016/j.jlb.2025.100303","DOIUrl":"10.1016/j.jlb.2025.100303","url":null,"abstract":"<div><div>Lung adenocarcinoma is a subtype of NSCLC that is often associated with poor prognosis. We present a case of metastatic lung adenocarcinoma in which comprehensive genomic profiling using both solid and liquid biopsies was employed to monitor disease progression and guide targeted therapy decisions. An initial liquid biopsy detected <em>ROS1-CCDC6</em> gene fusion, and the patient was started on Crizotinib. Following disease progression, further genomic profiling using both solid tissue and cfDNA revealed the presence of a previously undetected classical mutation <em>EGFR</em> exon 21, p. L858R. Consequently, the treatment was adjusted to include both Crizotinib and Gefitinib. A 6-month follow-up showed relapse and extensive metastasis. A repeat liquid biopsy identified a newly acquired <em>TP53</em> mutation (exon 7, p.R248Q) in addition to the persistent <em>EGFR</em> mutation. Restarting the targeted therapy led to complete metabolic resolution of the disease. This case highlights the utility of liquid biopsy when tissue biopsy is not feasible and underscores the importance of integrating both solid and liquid genomic data to capture a more comprehensive mutational landscape, including low-frequency or emerging variants, ultimately enabling more effective, individualized treatment strategies.</div></div>","PeriodicalId":101235,"journal":{"name":"The Journal of Liquid Biopsy","volume":"8 ","pages":"Article 100303"},"PeriodicalIF":0.0,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144240944","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cancer in a Drop: Liquid biopsy insights from AACR 2025","authors":"Shivahamy Maheswaran ,&nbsp;Roberto Borea ,&nbsp;Diego de Miguel Perez ,&nbsp;Pasquale Pisapia ,&nbsp;Nadia Ghazali ,&nbsp;Canio Martinelli ,&nbsp;Pavel Stejskal ,&nbsp;Carolina Reduzzi","doi":"10.1016/j.jlb.2025.100304","DOIUrl":"10.1016/j.jlb.2025.100304","url":null,"abstract":"","PeriodicalId":101235,"journal":{"name":"The Journal of Liquid Biopsy","volume":"8 ","pages":"Article 100304"},"PeriodicalIF":0.0,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144289016","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Corrigendum to “PP01.28 Expanding the Impact of Liquid Biopsies in patients with newly diagnosed advanced lung cancer (Expandinglung)” [J Liq Biopsy 5 (2024) e1–e32 100206]","authors":"Nadia Ghazali ,&nbsp;Mary R. Rabey ,&nbsp;Lisa Le ,&nbsp;M. Safi ,&nbsp;Christopher Pettengell ,&nbsp;Michelle Nguyen ,&nbsp;Maxym Muzychenko ,&nbsp;Steven Aviv ,&nbsp;Sachin Paleja ,&nbsp;Noor Ghumman ,&nbsp;Amina Ahmed ,&nbsp;Penelope A. Bradbury ,&nbsp;Frances A. Shepherd ,&nbsp;Geoffrey Liu ,&nbsp;Adrian G. Sacher ,&nbsp;Peter J.B. Sabatini ,&nbsp;Tracy L. Stockley ,&nbsp;Natasha B. Leighl","doi":"10.1016/j.jlb.2025.100302","DOIUrl":"10.1016/j.jlb.2025.100302","url":null,"abstract":"","PeriodicalId":101235,"journal":{"name":"The Journal of Liquid Biopsy","volume":"8 ","pages":"Article 100302"},"PeriodicalIF":0.0,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144270400","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The impact of liquid biopsy in breast cancer: Redefining the landscape of non-invasive precision oncology","authors":"Shaivy Malik,&nbsp;Sufian Zaheer","doi":"10.1016/j.jlb.2025.100299","DOIUrl":"10.1016/j.jlb.2025.100299","url":null,"abstract":"<div><div>Breast cancer (BC) remains a leading cause of morbidity and mortality among women worldwide, necessitating the development of innovative diagnostic and monitoring strategies. Liquid biopsy (LB), a minimally invasive approach that analyzes circulating tumor cells (CTCs), cell-free DNA (cfDNA), circulating tumor DNA (ctDNA), extracellular vesicles (EVs), and other tumor-derived biomarkers in body fluids, has emerged as a transformative tool in BC management. This review comprehensively explores the role of LB in early detection, disease monitoring, treatment stratification, and resistance surveillance in BC. We discuss the latest advancements in LB technologies, including next-generation sequencing (NGS), digital PCR, and single-cell analysis, highlighting their sensitivity and specificity. Additionally, we examine the clinical utility of LB in guiding personalized therapy, particularly in the context of hormone receptor-positive, HER2positive, and triple-negative BC subtypes. Despite its promise, several challenges, including standardization, validation, and integration into clinical practice, remain to be addressed. By summarizing current evidence and future directions, this review underscores the potential of LB to revolutionize BC diagnosis and treatment, paving the way for a more precise and dynamic approach to disease management.</div></div>","PeriodicalId":101235,"journal":{"name":"The Journal of Liquid Biopsy","volume":"8 ","pages":"Article 100299"},"PeriodicalIF":0.0,"publicationDate":"2025-05-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144167272","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Circulating tumor DNA profiling for non-invasive genomic analysis in Indian lung cancer patients: A real-world experience","authors":"Prerana Jha ,&nbsp;Rohit Mishra ,&nbsp;Asim Joshi ,&nbsp;Neha Sharma ,&nbsp;Minit Shah ,&nbsp;Govind Babu ,&nbsp;Amit Rauthan ,&nbsp;Sewanti Limaye ,&nbsp;Nandini Menon ,&nbsp;Venkataramanan Ramachandran ,&nbsp;Vanita Noronha ,&nbsp;Prashant Kumar ,&nbsp;Kumar Prabhash","doi":"10.1016/j.jlb.2025.100300","DOIUrl":"10.1016/j.jlb.2025.100300","url":null,"abstract":"<div><h3>Background</h3><div>Liquid biopsy assays are an important tool for non-invasive detection of genetic alterations, providing an effective alternative to traditional tissue biopsies. The study aimed to investigate the utility of ctDNA based next generation sequencing for clinical management of lung cancer patients from India.</div></div><div><h3>Methods</h3><div>We conducted ctDNA targeted sequencing on 425 lung cancer patients from India using 50 gene oncomine precision assay. The assay was validated employing 7 controls and 77 clinical samples, and the performance of the assay was evaluated. The concordance analysis with matched tissue biopsy samples was performed on 162 cases.</div></div><div><h3>Results</h3><div>Among the 425 lung cancer samples, 47 % harbored at least one mutation. <em>EGFR</em> was the most frequently altered gene (25.2 %), followed by <em>TP53</em> (19.8 %) and <em>KRAS</em> (4.5 %). Concordance with tissue biopsy data was 77 % for <em>EGFR</em>, 79 % for <em>TP53</em> and above 97 % for low frequency mutations. The assay demonstrated 100 % specificity and around 60 % sensitivity for the majority of clinically relevant genetic alterations including <em>EGFR, KRAS</em> and <em>BRAF</em>. Notably ERBB2 alterations were detected with 100 % sensitivity and specificity.</div></div><div><h3>Conclusion</h3><div>The ctDNA assay demonstrates high accuracy and specificity, for both prevalent and rare genetic alterations. While further advancements are needed to enhance sensitivity and routine clinical application, our ctDNA profiling assay offers a reliable alternative for detecting genetic alterations in lung cancer patients, with significant potential for clinical integration in the Indian healthcare context.</div></div>","PeriodicalId":101235,"journal":{"name":"The Journal of Liquid Biopsy","volume":"8 ","pages":"Article 100300"},"PeriodicalIF":0.0,"publicationDate":"2025-05-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144139044","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Analytical validation of Aspyre Clinical Test for Lung (Blood): A multiplexed PCR and pyrophosphorolysis-based assay for detecting actionable NSCLC variants in plasma cfDNA and cfRNA","authors":"Ryan Thomas Evans ,&nbsp;Katherine Elizabeth Knudsen ,&nbsp;Elizabeth Gillon-Zhang ,&nbsp;Julia Natalie Brown ,&nbsp;Candace King ,&nbsp;Mary Beth Rossi ,&nbsp;Cory Kiser ,&nbsp;James Alexander Schaffernoth ,&nbsp;Amanda Shull Green ,&nbsp;Ana-Luisa Silva ,&nbsp;Kristine von Bargen ,&nbsp;Justyna Malgorzata Mordaka ,&nbsp;Rebecca Natalie Palmer ,&nbsp;Alessandro Tomassini ,&nbsp;Alejandra Collazos ,&nbsp;Simonetta Andreazza ,&nbsp;Iyelola Turner ,&nbsp;Chau Ha Ho ,&nbsp;Dilyara Nugent ,&nbsp;Jinsy Jose ,&nbsp;Shari Brown","doi":"10.1016/j.jlb.2025.100298","DOIUrl":"10.1016/j.jlb.2025.100298","url":null,"abstract":"<div><h3>Background</h3><div>Liquid biopsy is an important non-invasive method of sampling the molecular profile of tumors for patients to access personalized oncology therapeutics but can be challenging. NGS-based methods require high sample quality, high sequencing depth and associated cost, with complex workflows, while PCR assays are limited in variant coverage. Aspyre Clinical Test for Lung® (Blood) is a simplified genomic profiling assay for NSCLC that targets 114 variants in 11 genes (<em>ALK, BRAF, EGFR, ERBB2, KRAS, RET, ROS1, MET &amp; NTRK1/2/3</em>) to robustly inform clinical management. The assay detects single nucleotide variants, insertions, deletions, gene fusions and exon skipping events from plasma-derived cfDNA and cfRNA simultaneously.</div></div><div><h3>Method</h3><div>Sensitivity, specificity, analytical accuracy and analytical precision at standard input levels (20 ng cfDNA and 42 ng cfRNA) were tested using a combination of contrived samples and extracts from clinical samples taken from both healthy volunteers and patients with NSCLC. The effects of potential interfering substances on assay performance were tested. Assay sensitivity and specificity were also assessed at lower sample input levels (5 ng cfDNA and 6 ng cfRNA).</div></div><div><h3>Results</h3><div>At standard input levels, median limits of detection were ≤0.25 % variant allele fraction for single nucleotide variants, ≤0.4 % variant allele fraction for insertions or deletions, ≤6 copies for gene fusions, and ≤100 copies <em>MET</em> exon 14 skipping events. The specificity from variant-free samples was 100 %. Tests of analytical accuracy yielded 100 % NPA and 94 % PPA between Aspyre Clinical Test for Lung (Blood) and either results from orthogonal NGS testing or expected outcomes of contrived samples. Results were 100 % replicable across multiple operators, reagent lots, days and equipment. At low input levels, median limits of detection were ≤0.8 % for single nucleotide variants and insertions/deletions, 6 copies for gene fusions and 100 copies for <em>MET</em> exon 14 skipping, with a false-positive rate of 0 %.</div></div><div><h3>Conclusions</h3><div>We present validation studies of Aspyre Clinical Test for Lung (Blood) using contrived and clinical samples. The technology is simple and fast, yet highly sensitive, specific, robust and reproducible with a turnaround time of two days. Aspyre Clinical Test for Lung (Blood) facilitates access to cost-effective, rapid, actionable molecular profiling of plasma for patients with NSCLC.</div></div>","PeriodicalId":101235,"journal":{"name":"The Journal of Liquid Biopsy","volume":"8 ","pages":"Article 100298"},"PeriodicalIF":0.0,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144134218","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}