Journal of Molecular Diagnostics最新文献

{"title":"Clinical Utility and Performance of Methylation-Specific Triplet-Primed PCR for Fragile X Syndrome Diagnosis","authors":"Areerat Hnoonual ,&nbsp;Wipawan Arunthong ,&nbsp;Oradawan Plong-on ,&nbsp;Pornsiri Sangmanee ,&nbsp;Pornprot Limprasert","doi":"10.1016/j.jmoldx.2025.10.001","DOIUrl":"10.1016/j.jmoldx.2025.10.001","url":null,"abstract":"<div><div>Fragile X syndrome (FXS) is the most common cause of intellectual disability. It is usually caused by the expansion of the CGG trinucleotide repeat (&gt;200 repeats) in <em>FMR1</em>, resulting in DNA hypermethylation and gene silencing. Conventional FXS diagnosis is based on a combination of PCR and Southern blot (SB) analysis, which is technically challenging and labor-intensive. Methylation-specific triplet-primed PCR (msTP-PCR) has been proposed to overcome these limitations in detecting <em>FMR1</em> expansion and methylation status. However, its performance in FXS diagnosis in clinical laboratory settings has not been extensively studied. An msTP-PCR assay was validated, implemented, and compared versus the conventional diagnostic protocol in clinical samples. A total of 420 clinical samples (339 male subjects and 81 female subjects) previously genotyped for <em>FMR1</em> CGG repeat expansion using reference methods (PCR-based methods and/or SB analysis) were investigated using msTP-PCR. The results showed high concordance with the reference results for allele categorization, repeat number correlation, and methylation status. However, discordant results were observed in rare cases of female patients with complex mosaic normal, premutation, and full mutation alleles, which need further confirmation via SB analysis. Nonetheless, these results confirm that the msTP-PCR method is a useful alternative technique for FXS diagnosis and prenatal testing, as it is rapid, reliable, cost-effective, and can potentially reduce the requirement for SB analysis.</div></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"28 1","pages":"Pages 112-123"},"PeriodicalIF":3.4,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145410638","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 Comprehension of the Pathogenicity of Splicing Variants","authors":"Yan Xu ,&nbsp;Bin Hu ,&nbsp;Xu Han ,&nbsp;Yiyao Chen ,&nbsp;Jingqi Lin ,&nbsp;Yanlin Wang ,&nbsp;Jian Wang ,&nbsp;Niu Li ,&nbsp;Shuyuan Li","doi":"10.1016/j.jmoldx.2025.10.006","DOIUrl":"10.1016/j.jmoldx.2025.10.006","url":null,"abstract":"<div><div>Despite the availability of various <em>in silico</em> prediction tools, accurately assessing the pathogenicity of splice-region variants remains limited. In this study, 18 splice-region variants of uncertain significance (VUSs) were functionally characterized using either <em>in vitro</em> (minigene) or <em>in vivo</em> (RT-PCR) experiments. The impacts of in-frame mutants on the indicated protein were further analyzed by modeling the three-dimensional protein structures. The accuracy of different <em>in silico</em> prediction tools (SpliceAI, varSEAK, and Splicing Prediction Pipeline) was also compared. The 18 VUSs included 3 canonical and 15 noncanonical splicing variants. Splicing abnormalities were observed in 16 variants (88.9%), with exon skipping (33.3%, 6/18) being the most prevalent event. Among them, 11 variants were predicted to generate premature termination codons, 3 caused in-frame alterations, and 2 resulted in both outcomes. Structural modeling revealed significant disruption of protein secondary structure in four of five in-frame alterations. Integrating functional and structural evidence, 15 VUSs (83.3%) were reclassified as pathogenic/likely pathogenic, enabling definitive diagnoses and informed reproductive decisions for affected families. Among the <em>in silico</em> tools, SpliceAI demonstrated the highest accuracy in predicting splicing anomalies; however, all tools showed decreased accuracy in analyzing specific splicing patterns. This study establishes an integrated workflow for assessing the pathogenicity of splicing VUSs in clinical diagnostics, offering a practical approach to overcome this critical diagnostic challenge.</div></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"28 1","pages":"Pages 75-84"},"PeriodicalIF":3.4,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145426677","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":"Clinical Validation of Duoseq, a Novel Assay for Clinical DNA and RNA Sequencing","authors":"Elizabeth Thacker ,&nbsp;Lanie Happ ,&nbsp;Magdalena Czader ,&nbsp;Lin Wang ,&nbsp;Christopher Giauque ,&nbsp;Jennifer Shingleton ,&nbsp;Cassandra Love ,&nbsp;Clay Parker ,&nbsp;Devang Thakkar ,&nbsp;Matt Sperling ,&nbsp;Kemin Xu ,&nbsp;Kenneth Ofori ,&nbsp;Andrea Vrydaghs ,&nbsp;Jen Loy-Flynn ,&nbsp;Robert Erno ,&nbsp;Callie Barker ,&nbsp;Jordan Bouldin ,&nbsp;Eric D. Hsi ,&nbsp;Sandeep S. Dave","doi":"10.1016/j.jmoldx.2025.10.003","DOIUrl":"10.1016/j.jmoldx.2025.10.003","url":null,"abstract":"<div><div>Next-generation sequencing (NGS) has become integral to the clinical workup of blood cancers. However, there remain barriers to implementing clinical NGS, including the separate workflows required for DNA and RNA sequencing, complex bioinformatics analyses, and long turnaround times. Duoseq has been developed as a comprehensive, kitted solution to these barriers and implemented as a genomic profiling tool for blood cancers to simultaneously detect single-nucleotide variants (SNVs), small insertions/deletions (indels), and structural variants (SVs; translocations and fusions). Additionally, Duoseq can evaluate numerous other blood cancer diagnostic markers, including gene expression, copy number alterations, cell of origin, oncogenic viral status (eg, Epstein-Barr virus), B-/T-cell receptor clonality, Ig heavy chain mutational status, and diffuse large B-cell lymphoma subtypes. Analytical and clinical validation of Duoseq was performed in parallel at two clinical institutions. Limit of detection was confirmed as 5% variant allele frequency for SNVs, 10% variant allele frequency for indels, and ≥20% tumor purity for SVs. Repeatability studies showed &gt;99% intrarun and interrun positive predictive value and &gt;96% percentage positive agreement. Duoseq was run on formalin-fixed, paraffin-embedded biopsy specimens (<em>N</em> = 197), using NGS and/or fluorescence <em>in situ</em> hybridization as orthogonal comparison. SNVs, indels, and SVs achieved accuracy of &gt;95%. These results establish Duoseq as a genomic profiling tool for blood cancers that can enable laboratories to provide critical diagnostic information in a time- and cost-effective manner.</div></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"28 1","pages":"Pages 85-100"},"PeriodicalIF":3.4,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145410798","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":"Clinical Performance of Tissue- and Plasma-Based Diagnostic Assays in Identifying Homologous Recombination Repair Gene Alterations in Patients with Metastatic Castration-Resistant Prostate Cancer following Treatment with Niraparib with Abiraterone Acetate Plus Prednisone (Niraparib + AAP)","authors":"Yaji Xu ,&nbsp;Usha Singh ,&nbsp;Katherine Bell ,&nbsp;Lesley Farrington ,&nbsp;Karen Urtishak ,&nbsp;Michael Gormley ,&nbsp;Won Kim ,&nbsp;Jenny Zhang ,&nbsp;Kristy Potts ,&nbsp;Songbai Wang","doi":"10.1016/j.jmoldx.2025.10.004","DOIUrl":"10.1016/j.jmoldx.2025.10.004","url":null,"abstract":"<div><div>This retrospective study was designed to demonstrate the clinical utility of two diagnostic tests, the FoundationOneCDx (F1CDx) and Exact Sciences Resolution homologous recombination deficiency (HRD; Resolution HRD assay) as clinical trial–enrollment assays to identify patients with metastatic castration-resistant prostate cancer harboring homologous recombination repair (HRR) gene alterations. Tumor tissue and plasma collected from patients with metastatic castration-resistant prostate cancer in the phase 3 MAGNITUDE study were tested using the F1CDx tissue assay and/or the Resolution HRD plasma assay. Patients with HRR alterations were randomized (1:1) to receive niraparib (NIRA) and abiraterone acetate (AA) + prednisone (NIRA + AAP) or placebo and AAP (PBO + AAP; NCT03748641). Efficacy was based on radiographic progression-free survival (primary end point), time tosymptomatic progression, time to cytotoxic chemotherapy, and overall survival as secondary end points (interim analysis 1). Of 423 HRR patients, 291 (68.8%) were HRR positive (HRR⁺) by F1CDx [<em>BRCA</em>^<em>+</em>: 162 (38.2%)], and 38 (8.9%) were HRR negative by F1CDx but HRR⁺ by Resolution HRD assay. Also, 277 of 423 (65.5%) were HRR⁺ by Resolution HRD assay [<em>BRCA</em>⁺: 150 (35.5%)], and 124 (29.3%) were HRR negative by Resolution HRD assay but HRR⁺ by F1CDx assay. Clinically meaningful benefits for all end points were comparable for <em>BRCA</em>^<em>+</em> and HRR⁺ patients detected by either tissue or plasma assays. These results demonstrated the clinical utility of both tissue and plasma assays in identifying patients for NIRA + AAP treatment.</div></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"28 1","pages":"Pages 101-111"},"PeriodicalIF":3.4,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145410655","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":"Validation of a Modular Gene Expression Assay for Risk Stratification and Subtyping Lymphomas","authors":"Peter J.B. Sabatini ,&nbsp;Josh Bridgers ,&nbsp;Shujun Huang ,&nbsp;Tong Zhang ,&nbsp;Clare Sheen ,&nbsp;Tracy Stockley ,&nbsp;Robert Kridel ,&nbsp;Ian Bosdet ,&nbsp;Marco A. Marra ,&nbsp;Christian Steidl ,&nbsp;David W. Scott ,&nbsp;Aly Karsan","doi":"10.1016/j.jmoldx.2025.09.004","DOIUrl":"10.1016/j.jmoldx.2025.09.004","url":null,"abstract":"<div><div>Gene expression signatures are important for classifying lymphoid malignancies, although routine diagnostic workflows predominantly use immunohistochemical staining and fluorescence <em>in situ</em> hybridization. These traditional methods are labor intensive and may misclassify the underlying oncogenic signatures, leading to inaccurate prognostication. To address this issue, an RNA expression panel was developed, the Lymphoma Expression Analysis (LExA120) 120 gene expression panel, using the NanoString platform for rapid, modular analysis of various lymphoma subtypes. The LExA120 panel targets 95 genes and 25 housekeeping genes to evaluate aggressive B-cell lymphomas, including: diffuse large B-cell lymphoma cell-of-origin, dark zone, and primary mediastinal large B-cell lymphoma signatures; Epstein-Barr virus (EBV) status; and a classical Hodgkin lymphoma posttransplant risk. Fifty-four formalin-fixed, paraffin-embedded tissue samples were tested with known diagnoses and 51 samples with known EBV status. The panel showed high concordance with previously validated methods according to Pearson correlation coefficients of the signature scores. The assay also displayed high reproducibility in repeated tests and across different clinical laboratories. This study confirmed the panel’s ability to stratify EBV-positive and EBV-negative lymphomas with high diagnostic certainty. Although EBER <em>in situ</em> hybridization confirmation was needed in approximately 12% of cases, synergizing with traditional techniques may facilitate more rapid and cost-effective diagnoses. The LExA120 panel offers a multiplexed approach to lymphoma classification, enhancing the efficiency and accuracy for subtyping lymphomas.</div></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"28 1","pages":"Pages 1-7"},"PeriodicalIF":3.4,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145281585","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":"Preanalytical Histology Review Improves POLE Mutation Detection in Endometrial Carcinomas","authors":"Hehua Huang,&nbsp;Sara Hartinger,&nbsp;Rachel L.P. Geisick,&nbsp;Chandler Ho,&nbsp;Fei Dong","doi":"10.1016/j.jmoldx.2025.09.008","DOIUrl":"10.1016/j.jmoldx.2025.09.008","url":null,"abstract":"<div><div>Preanalytical tissue assessment is an important step in cancer molecular testing; however, its impact on molecular test results has not been systematically evaluated. This study describes a quality-improvement project in which routine histology review was implemented at a US molecular diagnostics laboratory. The effects of implementation on laboratory compliance and the analytical performance of a targeted <em>POLE</em> assay were measured as changes in tumor cellularity documentation, tumor sample enrichment (in samples with &lt;40% tumor cellularity), <em>POLE</em> mutation rate, tumor signal intensity, and repeat-testing rate. Endometrial carcinoma samples (<em>N</em> = 1752) and tested for <em>POLE</em> mutations using a multiplex PCR assay. <em>POLE</em> mutation rates were 6.3% and 5.0% before and after intervention, respectively (<em>P</em> = 0.25), with the mutations most commonly detected being p.Pro286Arg (47%) and p.Val411Leu (21%). Documentation of tumor cellularity increased from 29% to 100%, and the rate of tumor enrichment increased from 1.4% to 31.5% (both, <em>P</em> &lt; 0.0001). Mutation signal intensity increased from 0.32 to 0.58, and the repeat-testing rate decreased from 8.8% to 2.3% (<em>P</em> = 0.004 and &lt;0.0001, respectively). Systematic preanalytical histology review was associated with improved analytical performance of a targeted <em>POLE</em> assay, accompanied by compliance in tumor cellularity documentation, increased tumor enrichment, and decreased repeated testing, supporting preanalytical assessment in improving somatic mutation detection in pathology specimens with low tumor content.</div></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"28 1","pages":"Pages 22-26"},"PeriodicalIF":3.4,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145330820","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 Splice-Altering Variants in Hematologic Malignancies Using Targeted RNA-Sequencing Data","authors":"Muneeza Maqsood ,&nbsp;John Toubia ,&nbsp;Carol Wadham ,&nbsp;Naranie Shanmuganathan ,&nbsp;Nur Hezrin Shahrin ,&nbsp;Adelina Fernandes ,&nbsp;Joe McConnell ,&nbsp;Verity Saunders ,&nbsp;Dominik Kaczorowski ,&nbsp;Rosalie R. Kenyon ,&nbsp;Ming Lin ,&nbsp;Timothy P. Hughes ,&nbsp;Chung Hoow Kok ,&nbsp;Susan Branford","doi":"10.1016/j.jmoldx.2025.09.003","DOIUrl":"10.1016/j.jmoldx.2025.09.003","url":null,"abstract":"<div><div>RNA-based targeted sequencing aids the detection of several types of variants in hematologic and other malignancies, including splice-altering variants. However, accurately identifying clinically relevant mis-splicing events remains challenging because of the inherent complexity of the human transcriptome and the high prevalence of false-positive splice junctions in deep RNA-sequencing data. To address these challenges, SpliceChaser and BreakChaser were developed, which are bioinformatics tools designed to enhance the detection and characterization of relevant splice-altering events. SpliceChaser improves the identification of clinically relevant atypical splicing by analyzing read length diversity within flanking sequences of the mapped reads around the splice junctions. BreakChaser processes soft-clipped sequences and alignment anomalies to enhance the detection of targeted deletion breakpoints associated with atypical splice isoforms generated from intrachromosomal gene deletions. These tools were developed and validated using a cohort of &gt;1400 RNA-sequencing samples from patients with chronic myeloid leukemia. Collectively, SpliceChaser and BreakChaser achieved a positive percentage agreement of 98% and a positive predictive value of 91% for the detection of clinically relevant atypical splice-altering variants or gene deletions in the targeted regions. By integrating splicing and breakpoint detection with robust filtering strategies, these tools facilitate precise identification of clinically relevant variants, paving the way for improved diagnostics and therapeutic strategies in chronic myeloid leukemia and other malignancies.</div></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"28 1","pages":"Pages 39-52"},"PeriodicalIF":3.4,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145281565","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":"Characterization of Large Genomic Rearrangements in BRCA1 and BRCA2 Genes in a Chinese High-Risk Cohort","authors":"Ava Kwong ,&nbsp;Cecilia Y.S. Ho ,&nbsp;Henry C.M. Leung ,&nbsp;Amy W.S. Leung ,&nbsp;Chun Hang Au ,&nbsp;Edmond S.K. Ma","doi":"10.1016/j.jmoldx.2025.09.005","DOIUrl":"10.1016/j.jmoldx.2025.09.005","url":null,"abstract":"<div><div>Large genomic rearrangements (LGRs) account for at least 10% of the mutations in <em>BRCA1</em> and 5% of <em>BRCA2</em> mutations in outbred families with hereditary breast and ovarian cancer. A total of 21 probands with breast cancer who carried <em>BRCA1</em> or <em>BRCA2</em> LGRs were identified from a cohort of 4678 Chinese patients. There was a total of 13 <em>BRCA1</em> LGR carriers and 8 <em>BRCA2</em> LGR carriers, including 12 large genomic deletions and 1 duplication. Ten and three specific breakpoints from <em>BRCA1</em> and <em>BRCA2</em>, respectively, were identified by either whole-genome sequencing by nanopore sequencing or long-range PCR. Five of these LGRs were recurrent LGRs. Three LGRs were novel founder LGRs in the southeast Chinese population. Chinese LGR carriers exhibited clinical phenotypes that were generally similar to those of non-LGR mutation carriers. However, there was a notable tendency for triple-negative breast cancer to be more prevalent among Chinese LGR carriers (<em>P</em> = 0.007), largely because of the predominance of <em>BRCA1</em> mutations. This suggests a potential association that warrants further investigation.</div></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"28 1","pages":"Pages 8-21"},"PeriodicalIF":3.4,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145801857","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":"Comparative Analysis of TP53-Mutated Tumor DNA in Saliva and Plasma of Patients with Head and Neck Squamous Cell Carcinoma","authors":"Liyona Kampel ,&nbsp;Shlomo Tsuriel ,&nbsp;Leonor L. Trejo ,&nbsp;Yaniv Hadi ,&nbsp;Gilad Horowitz ,&nbsp;Anton Warshavsky ,&nbsp;Dov Hershkovitz ,&nbsp;Nidal Muhanna","doi":"10.1016/j.jmoldx.2025.10.002","DOIUrl":"10.1016/j.jmoldx.2025.10.002","url":null,"abstract":"<div><div>Liquid biopsy offers a promising noninvasive alternative for tissue sampling in solid cancers. Saliva, an easily accessible biofluid, can harbor tumor DNA, yet its clinical utility compared with plasma circulating tumor DNA (ctDNA) in head and neck squamous cell carcinoma (HNSCC) remains uncertain. Tumor samples from patients with HNSCC underwent next-generation sequencing to identify <em>TP53</em> mutations. Matched plasma and saliva cell-free DNA (cfDNA) samples were analyzed for the presence of tumor-specific mutations. Pathologic features and survival data were evaluated in relation to mutation detectability in biofluids. Overall, <em>TP53</em> mutations were detected in 64 of 85 tumors (75%). Liquid biopsy analysis included 36 plasma and 21 saliva samples from 40 patients. Plasma ctDNA was detected in 68% of oral cavity squamous cell carcinoma compared with 29% of laryngeal cancer cases, and it was associated with nodal metastases (<em>P</em> = 0.034). Detection of ctDNA showed a trend toward worse progression-free survival (37.4 versus 68.5 months; <em>P</em> = 0.134). Tumor mutation was identified in saliva cfDNA in 57% of cases, irrespective of disease stage or presence of regional metastases. Plasma ctDNA emerged as a potential prognostic marker in HNSCC, whereas mutated saliva cfDNA, although frequently detectable, lacked prognostic value or correlation with adverse pathologic features. Further research is warranted to elucidate the mechanisms governing tumor DNA shedding into saliva and to define its clinical applications.</div></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"28 1","pages":"Pages 124-131"},"PeriodicalIF":3.4,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145410745","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 Targeted LC-MS/MS–Based Quantitative Assay for Detecting Plasma Factor XIII A/B Subunit Deficiency","authors":"Jinyong Kim ,&nbsp;Shilpa Venkataraman ,&nbsp;Sandip N. Chavan ,&nbsp;Ramesh Bokka ,&nbsp;Julie I. Tange ,&nbsp;Rachel R. Leger ,&nbsp;Rajiv K. Pruthi ,&nbsp;Dong Chen ,&nbsp;Jansen N. Seheult ,&nbsp;Akhilesh Pandey","doi":"10.1016/j.jmoldx.2025.09.010","DOIUrl":"10.1016/j.jmoldx.2025.09.010","url":null,"abstract":"<div><div>Factor XIII (FXIII) is a heterotetramer that plays a crucial role in the coagulation cascade, facilitating fibrin crosslinking to stabilize clots. Congenital or acquired deficiency of the FXIII A or B subunits can lead to prolonged bleeding. Existing enzyme-linked immunosorbent assay–based and latex agglutination assays for measuring FXIII are limited to detecting only the FXIIIA subunit of the complex. In this study, a targeted liquid chromatography–tandem mass spectrometry (LC-MS/MS) assay was developed and validated to accurately quantify both FXIIIA and FXIIIB subunits in plasma. Peptides measured by this targeted assay were rigorously selected based on analytical robustness, with the method demonstrating excellent sensitivity (limit of detection, 1.56 to 3.67 mU/mL), linearity (<em>r</em>² &gt; 0.998), and precision (CV, &lt;10%). Comparison with the conventional assay based on ammonia release demonstrated a strong correlation (<em>r</em> = 0.983) and agreement (Cohen κ = 0.846) for FXIIIA. On analysis of 98 clinical plasma samples, the assay accurately identified cases with FXIII deficiency based on significantly reduced FXIIIA with varying alterations in FXIIIB levels. These results establish the clinical utility of the LC-MS/MS assay, highlight its potential for improving diagnostic accuracy for FXIII deficiency, and lay the foundation for future research on FXIII dynamics in pathologic states.</div></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"28 1","pages":"Pages 64-74"},"PeriodicalIF":3.4,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145426607","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}