Journal of Molecular Diagnostics最新文献

{"title":"Validation of the SOPHiA DDM HRD Solution as a Companion Diagnostic for Poly (ADP-Ribose) Polymerase Inhibitor Access in Australia","authors":"Andrew P. Fellowes ,&nbsp;David Y.H. Choong ,&nbsp;Christopher R. McEvoy ,&nbsp;Roxane A. Legaie ,&nbsp;Anthony H. Bell ,&nbsp;Stephen B. Fox","doi":"10.1016/j.jmoldx.2025.12.002","DOIUrl":"10.1016/j.jmoldx.2025.12.002","url":null,"abstract":"<div><div>In this multilaboratory validation study of 145 ovarian cancer samples, the SOPHiA DDM HRD Solution was compared with the regulatory-approved Myriad myChoice HRD assay to assess clinical comparability for class 3 in-house <em>in vitro</em> diagnostic medical device companion diagnostic use. BRCA (<em>BRCA1</em> and <em>BRCA</em><em>2</em>) mutation status showed 100% concordance, and genomic instability (GI) measurements demonstrated strong linear agreement, absence of bias, and high analytical precision. Receiver operating characteristic analysis suggested a threshold adjustment from 0 to –1.5, improving overall accuracy to 91.2% when combined with BRCA mutation status to assign homologous recombination deficiency (HRD) status. Approximately 6% of samples were excluded because of inconclusive results, whereas GI classification discordance was concentrated near the clinical threshold. Neither inconclusiveness nor discordance was associated with sample-related factors. These findings indicate that the SOPHiA HRD assay can provide results broadly interchangeable with Myriad myChoice, although caution is warranted when assigning HRD status to borderline GI values.</div></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"28 3","pages":"Pages 294-307"},"PeriodicalIF":3.4,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145844189","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Prediction of Lung Metastasis in Breast Cancer Patients Using Machine Learning Classifiers","authors":"Thanh Dat Nguyen ,&nbsp;Quynh-Mai Thi Nguyen ,&nbsp;Tuong Van Nguyen ,&nbsp;Phuong Thi Bui ,&nbsp;Kim Nhuong Thi Nguyen ,&nbsp;Minh Nam Nguyen","doi":"10.1016/j.jmoldx.2025.10.010","DOIUrl":"10.1016/j.jmoldx.2025.10.010","url":null,"abstract":"<div><div>Breast cancer is the most common cancer among women, and metastasis to the lung is associated with poor prognosis. Reliable biomarkers for predicting lung metastasis are urgently needed to improve early detection and clinical decision-making. This study used microarray data sets comprising gene expression profiles and clinical data from primary breast cancer patients who were followed up for lung metastasis outcomes. High-throughput screening combined with Venn diagram analysis was used to identify common candidate probes, and the least absolute shrinkage and selection operator method were used to select 11 genes for model development. Logistic regression was used to construct predictive models, and the final risk signature consisted of 10 candidate genes (<em>CDK19, GLUD1, GTPBP4, HLCS, HYI, KCND3, MAP2K1, NMUR1, PRKD3</em>, and <em>SLC16A3</em>). The model achieved strong performance in training and validation cohorts (areas under the curve &gt;0.87) and generalized to the independent METABRIC data set (area under the curve = 0.706). Subset analyses restricted to patients with early-stage disease confirmed that the signature retained predictive value. Kaplan-Meier analyses showed that patients with high-risk scores had shorter lung metastasis–free survival, recurrence-free survival, and overall survival. Multivariate Cox analysis confirmed that the risk signature provided independent predictive information from clinical variables. In conclusion, the risk signature accurately identifies patients with breast cancer at risk of lung metastasis, enabling clinicians to better assess risk and tailor effective treatment strategies.</div></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"28 2","pages":"Pages 147-159"},"PeriodicalIF":3.4,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145642139","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Updates on the Clinical Epidemiology of HIV-1 Group O Strains in Cameroon and Potential Implications on Diagnosis and Treatment Strategies","authors":"Joseph Fokam ,&nbsp;Collins Ambe Chenwi ,&nbsp;Désiré Augustin Takou Komego ,&nbsp;Odile Estelle Grâce Beloumou Angong ,&nbsp;Sandrine Claire Djupsa Njdeyep ,&nbsp;Ezechiel Ngoufack Jagni Semengue ,&nbsp;Alex Durand Nka ,&nbsp;Aude Christelle Ka'e ,&nbsp;Vincent Kamaël Mekel ,&nbsp;Aurelie Minelle Kengni Ngueko ,&nbsp;Naomi-Karell Etame ,&nbsp;Rachel Audrey Nayang Mundo ,&nbsp;Samuel Martin Sosso ,&nbsp;Rachel Kamgaing ,&nbsp;Nadine Nguendjoung Fainguem ,&nbsp;Derrick Tambe Ayuk Ngwese ,&nbsp;Charles Fokunang ,&nbsp;Michel Carlos Tchouaket Tommo ,&nbsp;Joelle Bouba Pamen ,&nbsp;Alice Ketchaji ,&nbsp;Malachie Manaouda","doi":"10.1016/j.jmoldx.2025.10.008","DOIUrl":"10.1016/j.jmoldx.2025.10.008","url":null,"abstract":"<div><div>Cameroon is an epicenter of diverse HIV-1 strains, with diagnostic and management challenges. The objective herein was to update HIV-1 non-M prevalence and compare diagnostic performance of two- versus three-test algorithms. A facility-based study was conducted in February 2024 on 2207 HIV-1 clinical samples at the Chantal Biya International Reference Centre (Yaoundé, Cameroon). HIV-1 non-M were identified by molecular phylogeny. Rapid diagnostic tests (RDTs) used in the two-test (Determine and KHB colloidal gold) versus three-test (First Response, One Step, and KHB) algorithms were evaluated on non-M, with ACRO (HIV1/2 and p24) as independent RDT. No group N (0%) nor P (0%) was found, whereas nine group O were identified (0.4%; 95% CI, 0.2%–0.8%). For individuals harboring group O (mean age, 43 ± 12 years; 50% female), median (IQR): duration since HIV diagnosis was 627 (423 to 775) weeks; viremia, 12,385 (5340 to 72,682) copies/mL; and CD4 count, 52 (39 to 228) cells/mm³. One Step, KHB, and ACRO detected eight of eight group O (100%); First Response HIV1-2.0, seven of eight (87.5%); and Determine HIV1/2, six of eight (75%), <em>P</em> = 1.00. In this Cameroonian setting, HIV-1 group N and P are scarce, whereas group O remains low (&lt;1%). Transitioning from the two-test (75% performance) to the three-test algorithm (87.5% performance) could lead to improved diagnostic performance on currently circulating HIV-1 group O, calling for updates in RDTs to adapt to viral dynamics.</div></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"28 2","pages":"Pages 160-169"},"PeriodicalIF":3.4,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145566094","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Toward Comprehensive Detection of the SMN1/2 Genotypes","authors":"Aiko Iwata-Otsubo","doi":"10.1016/j.jmoldx.2025.11.001","DOIUrl":"10.1016/j.jmoldx.2025.11.001","url":null,"abstract":"","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"28 2","pages":"Pages 133-135"},"PeriodicalIF":3.4,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145650046","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Highly Sensitive Detection of Donor Chimerism by Next-Generation Sequencing","authors":"Eros Qama ,&nbsp;Abedul Haque ,&nbsp;Juan Du ,&nbsp;Abul K. Azad ,&nbsp;Rizwan Naeem ,&nbsp;Yanhua Wang ,&nbsp;Monika Paroder ,&nbsp;D. Yitzchak Goldstein ,&nbsp;David M. Loeb ,&nbsp;Adriana I. Colovai","doi":"10.1016/j.jmoldx.2025.11.004","DOIUrl":"10.1016/j.jmoldx.2025.11.004","url":null,"abstract":"<div><div>Donor chimerism analysis is used for monitoring engraftment status and risk of disease relapse following allogeneic stem cell transplantation. Recently developed assays using next-generation sequencing have demonstrated enhanced sensitivity and accuracy compared with standard capillary electrophoresis methods. This work presents validation results using One Lambda Devyser Chimerism assay, a next-generation sequencing–based test for monitoring donor chimerism. A total of 270 samples, including clinical and cell-line DNA, were tested. There was a high correlation between chimerism results obtained with One Lambda Devyser and short tandem repeat assays (<em>R</em>² = 0.998). Determination of the limit of blank, limit of detection, and limit of quantitation indicated that One Lambda Devyser Chimerism assay can reliably detect recipient DNA fractions as low as 0.1%. Analytical specificity was &gt;99.9%. Reproducibility, linearity, DNA library characteristics, and sequencing metrics are presented. The suitability of DNA markers was verified in a population predominantly African American and Hispanic, comprising 30 recipient/donor pairs. The average number of informative markers per pair was seven, with a lower representation (five markers) in related pairs. In conclusion, the results show that One Lambda Devyser Chimerism assay is a highly sensitive test for detecting donor chimerism in a diverse patient population. The assay performed remarkably well at low recipient concentrations, having the potential to detect early changes associated with disease relapse.</div></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"28 2","pages":"Pages 199-209"},"PeriodicalIF":3.4,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145650036","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optical Genome Mapping versus Whole-Genome Sequencing in the Clinical Diagnosis of Gynecologic Mesenchymal Tumors","authors":"Karin Wallander ,&nbsp;Yingbo Lin ,&nbsp;Vadym Ivanchuk ,&nbsp;Valeria Difilippo ,&nbsp;Venkatesh Chellappa ,&nbsp;Sarath K. Murugan ,&nbsp;Ingegerd Öfverholm ,&nbsp;Robert Bränström ,&nbsp;Karolin H. Nord ,&nbsp;Joseph Carlson ,&nbsp;Felix Haglund de Flon","doi":"10.1016/j.jmoldx.2025.11.003","DOIUrl":"10.1016/j.jmoldx.2025.11.003","url":null,"abstract":"<div><div>Optical genome mapping (OGM) enables high-resolution detection of structural variants (SVs) and copy number aberrations (CNAs) using ultralong DNA molecules and minimal bioinformatics processing. Its diagnostic utility in solid tumors remains underexplored. Whole-genome sequencing (WGS) offers comprehensive variant detection but is resource intensive. This study presents a technical benchmarking of OGM versus WGS for mesenchymal tumors of the gynecologic tract. Twenty-five uterine mesenchymal tumors were prospectively analyzed using matched WGS, transcriptome sequencing, and OGM. Detected SVs, CNAs, and fusion genes were compared across platforms. OGM identified structural driver events in 80% of cases and demonstrated high concordance with WGS for major CNAs and translocations. In select cases, OGM resolved complex rearrangements not clearly defined by WGS, including a <em>PLAG1</em>::<em>RERE</em> fusion and an embedded inversion in a <em>RAD51B</em>::<em>HMGA2</em> event. Conversely, WGS uniquely detected a truncating <em>NF1</em> translocation and a <em>TSC2</em>::<em>SENP3</em> fusion, both clinically significant. OGM is a technically robust platform for SV and CNA detection in mesenchymal tumors, and it may serve as an efficient alternative to sequencing-based cytogenomic approaches in selected clinical contexts, especially in tumors known to be driven by gross chromosomal rearrangements. WGS provides a comprehensive view of the cancer genome, suitable for tumors driven by single-nucleotide variants, SVs, and CNAs. The choice between platforms should be guided by clinical context, diagnostic needs, and available resources.</div></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"28 2","pages":"Pages 187-198"},"PeriodicalIF":3.4,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145650095","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enhanced Detection of EGFRvIII in Tumors","authors":"Nan Chen ,&nbsp;Jingjing Feng ,&nbsp;Dong Wan ,&nbsp;Dongbing Li ,&nbsp;Sheng Xiao","doi":"10.1016/j.jmoldx.2025.11.002","DOIUrl":"10.1016/j.jmoldx.2025.11.002","url":null,"abstract":"<div><div>Epidermal growth factor receptor variant (EGFRv)-III, a common oncogenic variant in glioblastoma and other solid tumors, results from an in-frame deletion of exons 2 to 7 from the <em>EGFR</em> gene. Detection of EGFRvIII is crucial for understanding tumor biology, guiding targeted therapies, and developing personalized treatment strategies. In this study, two detection approaches based on DNA next-generation sequencing—read depth (RD)-based and split read (SR)-based detection—were compared to evaluate their sensitivity and accuracy in identifying EGFRvIII. Thirty-one tumor samples, including glioblastoma and pancreatic adenocarcinoma, were analyzed using both methods. The SR-based method detected EGFRvIII in 20 of 31 samples, while the RD-based method identified it in only 12 samples (<em>P</em> &lt; 0.001), demonstrating that the SR-based method had significantly higher sensitivity. RNA next-generation sequencing confirmed EGFRvIII expression in most SR-positive cases. Additionally, the SR-based method identified multiple breakpoints in several tumors, revealing intratumor heterogeneity and the subclonal origins of EGFRvIII. The RD-based method was prone to false negatives, particularly in cases with high <em>EGFR</em> amplification or low tumor cell percentage, where copy number variations could be masked by background noise. The findings highlight the sensitivity and accuracy of SR-based detection in identifying EGFRvIII and capturing intratumor heterogeneity. SR-based analysis is recommended as the method for EGFRvIII detection in both clinical and research settings.</div></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"28 2","pages":"Pages 179-186"},"PeriodicalIF":3.4,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145650074","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A More Clinically Effective Long-Read Sequencing–Based Approach for Comprehensive Analysis of Spinal Muscular Atrophy","authors":"Shuyuan Li ,&nbsp;Bailing Liu ,&nbsp;Jingfan Zhang ,&nbsp;Ning Tang ,&nbsp;Renyi Hua ,&nbsp;Jinling Yang ,&nbsp;Xingling Huang ,&nbsp;Haoxian Li ,&nbsp;Aiping Mao ,&nbsp;Libao Chen ,&nbsp;Jiwei Huang ,&nbsp;Yanlin Wang","doi":"10.1016/j.jmoldx.2025.10.009","DOIUrl":"10.1016/j.jmoldx.2025.10.009","url":null,"abstract":"<div><div>Conventional methods for spinal muscular atrophy (SMA) screening have been challenging in detecting <em>SMN1/2</em> single-nucleotide variants (SNVs) and small insertions and deletions, <em>SMN1</em> 2 + 0 silent carrier, and the copy number (CN) of <em>SMN2</em>. To address these limitations, a long-read sequencing (LRS)-based approach termed comprehensive analysis of SMA 2 (CASMA2) was developed. CASMA2 was used to perform CN analysis by integrating Poisson distribution with an endogenous reference gene, the first such method developed for LRS platforms. The performance and clinical feasibility of CASMA2 were evaluated by using 414 retrospective peripheral blood samples and 303 prospective dried blood spot samples. CASMA2 displayed 100% accuracy in <em>SMN1</em>/<em>2</em> CN analysis and identified the SNVs/insertions and deletions in <em>SMN1/2</em>. CASMA2 also showed the capability of screening for the <em>SMN1</em> 2 + 0 silent carrier with family-trio haplotype analysis. It achieved a 99.0% (410 of 414) first-attempt success rate for long-term peripheral blood samples and a 98.7% (299 of 303) rate for dried blood spot samples. CASMA2 offers a clinically feasible, precise, and efficient method for SMA carrier and newborn screening.</div></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"28 2","pages":"Pages 136-146"},"PeriodicalIF":3.4,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145566110","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Functional Analysis of Four Splice-Site Variants, including a Novel Variant, on Antigen Expression and ABO Subgroup Formation","authors":"Huayue Yang ,&nbsp;Dan Long ,&nbsp;Hang Lei ,&nbsp;Liujun Guo ,&nbsp;Chengyan Gao ,&nbsp;Haoyu Hao ,&nbsp;Jiaming Li ,&nbsp;Xuefeng Wang ,&nbsp;Xi Wu ,&nbsp;Jing Dai ,&nbsp;Can Lou ,&nbsp;Xiaohong Cai","doi":"10.1016/j.jmoldx.2025.10.007","DOIUrl":"10.1016/j.jmoldx.2025.10.007","url":null,"abstract":"<div><div>Splice-site variants within the <em>ABO</em> gene have the potential to impair ABO glycosyltransferase biosynthesis, leading to decreased expression of A or B antigens on the surface of red blood cells. This study characterized how four intron 6 splice-site variants—three in the <em>A1</em> allele (c.374+4A&gt;T, c.374+4A&gt;G, and c.374+5G&gt;A) and one in the <em>ABO∗B.01</em> allele (c.374+2_374+3insT)—affect <em>ABO</em> pre-mRNA splicing. A combination of serologic, molecular genetic, bioinformatic, and minigene assays established a genotype-splicing-phenotype association model. Bioinformatics predictions showed that all variants impaired the recognition of the 5′ splice site. Specifically, the <em>A</em> allele variant c.374+5G&gt;A induced approximately 2.8% exon 6 retention, whereas the c.374+4A&gt;G (A_finn/A_weak) and c.374+4A&gt;T (A_weak) variants preserved approximately 6.9% and 10.2% functional transcripts, respectively. The novel <em>B</em> allele insertion variant c.374+2_374+3insT (B_el subtype) retained approximately 4.7% exon 6–containing transcripts, sufficient for trace B glycosyltransferase expression. Quantitative real-time PCR analysis confirmed that the transcriptional levels of the <em>ABO</em> gene in the AB_el subtype individual carrying the c.374+2_374+3insT variant were approximately 51.7% of those in the <em>ABO∗A1.02/ABO∗B.01</em> control. This study demonstrated that splice-site variants in the <em>ABO</em> gene reduce the abundance of functional transcripts via altered transcriptional regulation, which, in turn, leads to decreased ABO glycosyltransferase expression, ultimately resulting in weak antigen phenotypes of varying intensity.</div></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"28 2","pages":"Pages 170-178"},"PeriodicalIF":3.4,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145566107","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bridge Capture Permits Cost-Efficient, Rapid, and Sensitive Molecular Precision Diagnostics","authors":"Simona Adamusová ,&nbsp;Anttoni Korkiakoski ,&nbsp;Nea Laine ,&nbsp;Anna Musku ,&nbsp;Tuula Rantasalo ,&nbsp;Jorma Kim ,&nbsp;Juuso Blomster ,&nbsp;Jukka Laine ,&nbsp;Tatu Hirvonen ,&nbsp;Juha-Pekka Pursiheimo ,&nbsp;Manu Tamminen","doi":"10.1016/j.jmoldx.2025.09.006","DOIUrl":"10.1016/j.jmoldx.2025.09.006","url":null,"abstract":"<div><div>Liquid biopsies quantifying mutations in circulating tumor DNA by targeted next-generation sequencing have been gaining popularity. They are performed by various library preparation methods, each with distinct advantages and limitations. This work introduces Bridge Capture, a novel technology that goes beyond the advantages of market-leading liquid biopsy technologies, eliminating the need to compromise between scalability, cost-efficiency, sensitivity, or panel size. Twenty-four matched contrived colorectal biospecimens mimicking circulating tumor DNA were analyzed by Bridge Capture, Archer LIQUIDPlex, and AmpliSeq CHP version 2 for Illumina to compare variant allele frequency (VAF) detection. Bridge Capture was evaluated for sequencing depth requirement, interlaboratory reproducibility, automatization, and panel scalability. Of all methods, Bridge Capture detected the lowest VAF, and all VAFs strongly correlated with Archer LIQUIDPlex (<em>R</em>² = 0.995) and AmpliSeq CHPv2 for Illumina (<em>R</em>² = 0.988). Owing to its unique design, the Bridge Capture is compatible with the commonly used next-generation sequencing platforms and effectively uses sequencing capacity, permitting affordable and sensitive variant detection. The method demonstrated high reproducibility across independent laboratories and between automated and manual workflow. The panel size was increased by 300% and had negligible impact on performance and cross-reactivity of the probes, implying high multiplexing capabilities. Taken together, Bridge Capture is a cost-efficient, simple, rapid, and sensitive cancer diagnostics tool that has a potential to significantly improve the detection of mutations.</div></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"28 1","pages":"Pages 53-63"},"PeriodicalIF":3.4,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145330761","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}