Journal of Molecular Diagnostics最新文献

{"title":"FMR1 Protein Expression Correlates with Intelligence Quotient in Both Peripheral Blood Mononuclear Cells and Fibroblasts from Individuals with an FMR1 Mutation","authors":"Poonnada Jiraanont ,&nbsp;Marwa Zafarullah ,&nbsp;Noor Sulaiman ,&nbsp;Glenda M. Espinal ,&nbsp;Jamie L. Randol ,&nbsp;Blythe Durbin-Johnson ,&nbsp;Andrea Schneider ,&nbsp;Randi J. Hagerman ,&nbsp;Paul J. Hagerman ,&nbsp;Flora Tassone","doi":"10.1016/j.jmoldx.2024.02.007","DOIUrl":"10.1016/j.jmoldx.2024.02.007","url":null,"abstract":"<div><p>Fragile X syndrome (FXS) is the most common heritable form of intellectual disability and is caused by CGG repeat expansions exceeding 200 (full mutation). Such expansions lead to hypermethylation and transcriptional silencing of the fragile X messenger ribonucleoprotein 1 (<em>FMR1</em>) gene. As a consequence, little or no <em>FMR1</em> protein (FMRP) is produced; absence of the protein, which normally is responsible for neuronal development and maintenance, causes the syndrome. Previous studies have demonstrated the causal relationship between FMRP levels and cognitive abilities in peripheral blood mononuclear cells (PBMCs) and dermal fibroblast cell lines of patients with FXS. However, it is arguable whether PBMCs or fibroblasts would be the preferred surrogate for measuring molecular markers, particularly FMRP, to represent the cognitive impairment, a core symptom of FXS. To address this concern, CGG repeats, methylation status, <em>FMR1</em> mRNA, and FMRP levels were measured in both PBMCs and fibroblasts derived from 66 individuals. The findings indicated a strong association between <em>FMR1</em> mRNA expression levels and CGG repeat numbers in PBMCs of premutation males after correcting for methylation status. Moreover, FMRP expression levels from both PBMCs and fibroblasts of male participants with a hypermethylated full mutation and with mosaicism demonstrated significant association between the intelligence quotient levels and FMRP levels, suggesting that PBMCs may be preferable for FXS clinical studies, because of their greater accessibility.</p></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"26 6","pages":"Pages 498-509"},"PeriodicalIF":4.1,"publicationDate":"2024-03-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140208099","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":"Panel Comparative Analysis Tool","authors":"André Oszwald ,&nbsp;Lucia Zisser ,&nbsp;Eva Compérat ,&nbsp;Leonhard Müllauer","doi":"10.1016/j.jmoldx.2024.01.015","DOIUrl":"10.1016/j.jmoldx.2024.01.015","url":null,"abstract":"<div><p>Multigene next-generation sequencing (NGS) panels have become a routine diagnostic method in the contemporary practice of personalized medicine. To avoid inadequate test choice or interpretation, a detailed understanding of the precise panel target regions is required. However, the necessary bioinformatic expertise is not always available, and publicly accessible and easily interpretable analyses of target regions are scarce. To address this critical knowledge gap, we present the Panel Comparative Analysis Tool (PanelCAT), an open-source application to analyze, visualize, and compare NGS panel DNA target regions. PanelCAT uses Reference Sequence, ClinVar, and Catalogue of Somatic Mutations in Cancer mutation census databases to quantify the exon and mutation coverage of target regions and provides interactive graphical representations and search functions to inspect the results. We demonstrate the utility of PanelCAT by analyzing two large NGS panels (TruSight Oncology 500 and Human Pan Cancer Panel) to validate the advertised target genes, quantify targeted exons and mutations, and identify differences between panels. PanelCAT will enable institutions and researchers to catalog and visualize NGS panel target regions independent of the manufacturer, promote transparency of panel limitations, and share this information with employees and requisitioners.</p></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"26 5","pages":"Pages 423-429"},"PeriodicalIF":4.1,"publicationDate":"2024-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1525157824000564/pdfft?md5=3cbb26f6bddb209a195561c3ddb67d17&pid=1-s2.0-S1525157824000564-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140177474","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Deconvoluting the Complexity of Congenital Sideroblastic Anemias through Genetic and Functional Profiling","authors":"FNU Alnoor ,&nbsp;Robert S. Ohgami","doi":"10.1016/j.jmoldx.2024.03.002","DOIUrl":"10.1016/j.jmoldx.2024.03.002","url":null,"abstract":"","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"26 5","pages":"Pages 321-322"},"PeriodicalIF":4.1,"publicationDate":"2024-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1525157824000576/pdfft?md5=5035c823b78157604d948c036367e8ea&pid=1-s2.0-S1525157824000576-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140141051","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"DNA Reference Reagents for Genotyping RH Variants","authors":"Emilia Sippert ,&nbsp;Evgeniya Volkova ,&nbsp;Meagan Rippee-Brooks ,&nbsp;Gregory A. Denomme ,&nbsp;Willy A. Flegel ,&nbsp;Christine Lee ,&nbsp;Richardae Araojo ,&nbsp;Orieji Illoh ,&nbsp;Zhugong Liu ,&nbsp;Maria Rios","doi":"10.1016/j.jmoldx.2024.02.005","DOIUrl":"10.1016/j.jmoldx.2024.02.005","url":null,"abstract":"<div><p>Patients who carry Rhesus (RH) blood group variants may develop Rh alloantibodies requiring matched red blood cell transfusions. Serologic reagents for Rh variants often fail to specifically identify variant Rh antigens and are in limited supply. Therefore, red blood cell genotyping assays are essential for managing transfusions in patients with clinically relevant Rh variants. Well-characterized DNA reference reagents are needed to ensure quality and accuracy of the molecular tests. Eight lyophilized DNA reference reagents, representing 21 polymorphisms in <em>RHD</em> and <em>RHCE,</em> were produced from an existing repository of immortalized B-lymphoblastoid cell lines at the Center for Biologics Evaluation and Research/US Food and Drug Administration. The material was validated through an international collaborative study involving 17 laboratories that evaluated each DNA candidate using molecular assays to characterize <em>RHD</em> and <em>RHCE</em> alleles, including commercial platforms and laboratory-developed testing, such as Sanger sequencing, next-generation sequencing, and third-generation sequencing. The genotyping results showed 99.4% agreement with the expected results for the target <em>RH</em> polymorphisms and 87.9% for <em>RH</em> allele agreement. Most of the discordant <em>RH</em> alleles results were explained by a limited polymorphism coverage in some genotyping methods. Results of stability and accelerated degradation studies support the suitability of these reagents for use as reference standards. The collaborative study results demonstrate the qualification of these eight DNA reagents for use as reference standards for <em>RH</em> blood group genotyping assay development and analytical validation.</p></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"26 6","pages":"Pages 456-466"},"PeriodicalIF":4.1,"publicationDate":"2024-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140144491","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":"Evaluation of Pre-Analytical Variables for Human Papillomavirus Primary Screening from Self-Collected Vaginal Swabs","authors":"Michelle Qi ,&nbsp;Anissa R. Naranjo ,&nbsp;Abigail J. Duque ,&nbsp;Thomas S. Lorey ,&nbsp;Jeffrey M. Schapiro ,&nbsp;Betty J. Suh-Burgmann ,&nbsp;Michael Rummel ,&nbsp;Stephen J. Salipante ,&nbsp;Nicolas Wentzensen ,&nbsp;Dina N. Greene","doi":"10.1016/j.jmoldx.2024.02.006","DOIUrl":"10.1016/j.jmoldx.2024.02.006","url":null,"abstract":"<div><p>Human papillomavirus (HPV) primary screening is an effective approach to assessing cervical cancer risk. Self-collected vaginal swabs can expand testing access, but the data defining analytical performance criteria necessary for adoption of self-collected specimens are limited, especially for those occurring outside the clinic, where the swab remains dry during transport. Here, we evaluated the performance of self-collected vaginal swabs for HPV detection using the Cobas 6800. There was insignificant variability between swabs self-collected by the same individual (<em>n</em> = 15 participants collecting 5 swabs per participant), measured by amplification of HPV and human β-globin control DNA. Comparison of self-collected vaginal swab and provider-collected cervical samples (<em>n</em> = 144 pairs) proved highly concordant for HPV detection (total agreement = 90.3%; positive percentage agreement = 84.2%). There was no relationship between the number of dry storage days and amplification of HPV (<em>n</em> = 68; range, 4 to 41 days). Exposure of self-collected dry swabs to extreme summer and winter temperatures did not affect testing outcomes. A second internal control (RNase P) demonstrated that lack of amplification for β-globin from self-collected specimens was consistent with poor, but not absent, cellularity. These data suggest that self-collected vaginal samples enable accurate clinical HPV testing, and that extended ambient dry storage or exposure to extreme temperatures does not influence HPV detection. Furthermore, lack of β-globin amplification in HPV-negative samples accurately identified participants who required recollection.</p></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"26 6","pages":"Pages 487-497"},"PeriodicalIF":4.1,"publicationDate":"2024-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1525157824000552/pdfft?md5=2bd70eec725bee4de6ccd1056a8bc7f2&pid=1-s2.0-S1525157824000552-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140144492","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Diagnosis of Challenging Spinal Muscular Atrophy Cases with Long-Read Sequencing","authors":"Ningning Wang ,&nbsp;Kexin Jiao ,&nbsp;Jin He ,&nbsp;Bochen Zhu ,&nbsp;Nachuan Cheng ,&nbsp;Jian Sun ,&nbsp;Lan Chen ,&nbsp;Wanjin Chen ,&nbsp;Lingyun Gong ,&nbsp;Kai Qiao ,&nbsp;Jianying Xi ,&nbsp;Qihan Wu ,&nbsp;Chongbo Zhao ,&nbsp;Wenhua Zhu","doi":"10.1016/j.jmoldx.2024.02.004","DOIUrl":"10.1016/j.jmoldx.2024.02.004","url":null,"abstract":"<div><p>Spinal muscular atrophy (SMA) is an autosomal recessive neuromuscular disorder primarily caused by the deletion or mutation of the survival motor neuron 1 (<em>SMN1</em>) gene. This study assesses the diagnostic potential of long-read sequencing (LRS) in three patients with SMA. For Patient 1, who has a heterozygous <em>SMN1</em> deletion, LRS unveiled a missense mutation in <em>SMN1</em> exon 5. In Patient 2, an <em>Alu/Alu</em>-mediated rearrangement covering the <em>SMN1</em> promoter and exon 1 was identified through a blend of multiplex ligation-dependent probe amplification, LRS, and PCR across the breakpoint. The third patient, born to a consanguineous family, bore four copies of hybrid <em>SMN</em> genes. LRS determined the genomic structures, indicating two distinct hybrids of <em>SMN2</em> exon 7 and <em>SMN1</em> exon 8. However, a discrepancy was found between the <em>SMN1</em>/<em>SMN2</em> ratio interpretations by LRS (0:2) and multiplex ligation-dependent probe amplification (0:4), which suggested a limitation of LRS in SMA diagnosis. In conclusion, this newly adapted long PCR–based third-generation sequencing introduces an additional avenue for SMA diagnosis.</p></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"26 5","pages":"Pages 364-373"},"PeriodicalIF":4.1,"publicationDate":"2024-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140137422","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":"Assay Validation of Cell-Free DNA Shallow Whole-Genome Sequencing to Determine Tumor Fraction in Advanced Cancers","authors":"Micah Rickles-Young ,&nbsp;Gabriel Tinoco ,&nbsp;Junko Tsuji ,&nbsp;Sam Pollock ,&nbsp;Marcy Haynam ,&nbsp;Heather Lefebvre ,&nbsp;Kristyn Glover ,&nbsp;Dwight H. Owen ,&nbsp;Katharine A. Collier ,&nbsp;Gavin Ha ,&nbsp;Viktor A. Adalsteinsson ,&nbsp;Carrie Cibulskis ,&nbsp;Niall J. Lennon ,&nbsp;Daniel G. Stover","doi":"10.1016/j.jmoldx.2024.01.014","DOIUrl":"10.1016/j.jmoldx.2024.01.014","url":null,"abstract":"<div><p>Blood-based liquid biopsy is increasingly used in clinical care of patients with cancer, and fraction of tumor-derived DNA in circulation (tumor fraction; TFx) has demonstrated clinical validity across multiple cancer types. To determine TFx, shallow whole-genome sequencing of cell-free DNA (cfDNA) can be performed from a single blood sample, using an established computational pipeline (ichorCNA), without prior knowledge of tumor mutations, in a highly cost-effective manner. We describe assay validation of this approach to facilitate broad clinical application, including evaluation of assay sensitivity, precision, repeatability, reproducibility, pre-analytic factors, and DNA quality/quantity. Sensitivity to detect TFx of 3% (lower limit of detection) was 97.2% to 100% at 1× and 0.1× mean sequencing depth, respectively. Precision was demonstrated on distinct sequencing instruments (HiSeqX and NovaSeq) with no observable differences. The assay achieved prespecified 95% agreement of TFx across replicates of the same specimen (repeatability) and duplicate samples in different batches (reproducibility). Comparison of samples collected in EDTA and Streck tubes from single venipuncture in 23 patients demonstrated that EDTA or Streck tubes were comparable if processed within 8 hours. On the basis of a range of DNA inputs (1 to 50 ng), 20 ng cfDNA is the preferred input, with 5 ng minimum acceptable. Overall, this shallow whole-genome sequencing of cfDNA and ichorCNA approach offers sensitive, precise, and reproducible quantitation of TFx, facilitating assay application in clinical cancer care.</p></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"26 5","pages":"Pages 413-422"},"PeriodicalIF":4.1,"publicationDate":"2024-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1525157824000527/pdfft?md5=c9933d34a83a021ca1c718cd91c5c493&pid=1-s2.0-S1525157824000527-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140137421","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optical Genome Mapping for Comprehensive Cytogenetic Analysis of Soft-Tissue and Bone Tumors for Diagnostic Purposes","authors":"Jef Baelen ,&nbsp;Barbara Dewaele ,&nbsp;Maria Debiec-Rychter ,&nbsp;Raphael Sciot ,&nbsp;Patrick Schöffski ,&nbsp;Daphne Hompes ,&nbsp;Friedl Sinnaeve ,&nbsp;Hazem Wafa ,&nbsp;Isabelle Vanden Bempt","doi":"10.1016/j.jmoldx.2024.02.003","DOIUrl":"10.1016/j.jmoldx.2024.02.003","url":null,"abstract":"<div><p>Soft-tissue and bone tumors represent a heterogeneous group of tumors encompassing more than 100 histologic subtypes today. Identifying genetic aberrations increasingly is important in these tumors for accurate diagnosis. Although gene mutations typically are detected by second-generation sequencing, the identification of structural variants (SVs) and copy number alterations (CNAs) remains challenging and requires various cytogenetic techniques including karyotyping, fluorescence <em>in situ</em> hybridization, and arrays, each with important limitations. Optical Genome Mapping (OGM), a non–sequencing-based technique for high-resolution detection of SVs and CNAs, was applied in a retrospective series of diagnostic soft-tissue and bone tumor samples. Sample preparation was successful in 38 of 53 cases, with the highest success rate in nonadipocytic soft-tissue tumors (24 of 27 cases; 89%). In 32 of 35 cases carrying a diagnostic SV or CNA, OGM identified the aberration (91%), including a <em>POU2AF3::EWSR1</em> fusion in a round cell sarcoma and a translocation t(1;5)(p22;p15) in a myxoinflammatory fibroblastic sarcoma. Interestingly, OGM shed light on the genomic complexity underlying the various aberrations. In five samples, OGM showed that chains of rearrangements generated the diagnostic fusion, three of which involved chromoplexy. In addition, in nine samples, chromothripsis was causal to the formation of giant marker/ring/double-minute chromosomes. Finally, compared with standard-of-care cytogenetics, OGM revealed additional aberrations, requiring further investigation of their potential clinical relevance.</p></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"26 5","pages":"Pages 374-386"},"PeriodicalIF":4.1,"publicationDate":"2024-02-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S152515782400045X/pdfft?md5=6ccbe0e387e436a1141b6bbdb5dbbb66&pid=1-s2.0-S152515782400045X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139941109","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Machine Learning–Supported Diagnosis of Small Blue Round Cell Sarcomas Using Targeted RNA Sequencing","authors":"Lea D. Schlieben ,&nbsp;Maria Giulia Carta ,&nbsp;Evgeny A. Moskalev ,&nbsp;Robert Stöhr ,&nbsp;Markus Metzler ,&nbsp;Manuel Besendörfer ,&nbsp;Norbert Meidenbauer ,&nbsp;Sabine Semrau ,&nbsp;Rolf Janka ,&nbsp;Robert Grützmann ,&nbsp;Stefan Wiemann ,&nbsp;Arndt Hartmann ,&nbsp;Abbas Agaimy ,&nbsp;Florian Haller ,&nbsp;Fulvia Ferrazzi","doi":"10.1016/j.jmoldx.2024.02.002","DOIUrl":"10.1016/j.jmoldx.2024.02.002","url":null,"abstract":"<div><p>Small blue round cell sarcomas (SBRCSs) are a heterogeneous group of tumors with overlapping morphologic features but markedly varying prognosis. They are characterized by distinct chromosomal alterations, particularly rearrangements leading to gene fusions, whose detection currently represents the most reliable diagnostic marker. Ewing sarcomas are the most common SBRCSs, defined by gene fusions involving <em>EWSR1</em> and transcription factors of the ETS family, and the most frequent non–<em>EWSR1</em>-rearranged SBRCSs harbor a <em>CIC</em> rearrangement. Unfortunately, currently the identification of <em>CIC</em>::<em>DUX4</em> translocation events, the most common <em>CIC</em> rearrangement, is challenging. Here, we present a machine-learning approach to support SBRCS diagnosis that relies on gene expression profiles measured via targeted sequencing. The analyses on a curated cohort of 69 soft-tissue tumors showed markedly distinct expression patterns for SBRCS subgroups. A random forest classifier trained on Ewing sarcoma and <em>CIC</em>-rearranged cases predicted probabilities of being <em>CIC</em>-rearranged &gt;0.9 for <em>CIC</em>-rearranged–like sarcomas and &lt;0.6 for other SBRCSs. Testing on a retrospective cohort of 1335 routine diagnostic cases identified 15 candidate <em>CIC</em>-rearranged tumors with a probability &gt;0.75, all of which were supported by expert histopathologic reassessment. Furthermore, the multigene random forest classifier appeared advantageous over using high <em>ETV4</em> expression alone, previously proposed as a surrogate to identify <em>CIC</em> rearrangement. Taken together, the expression-based classifier can offer valuable support for SBRCS pathologic diagnosis.</p></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"26 5","pages":"Pages 387-398"},"PeriodicalIF":4.1,"publicationDate":"2024-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1525157824000461/pdfft?md5=1bdd20bb1eef2f4aac2f889d2553cfd5&pid=1-s2.0-S1525157824000461-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139921304","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Universal Digital High-Resolution Melt Analysis for the Diagnosis of Bacteremia","authors":"April Aralar ,&nbsp;Tyler Goshia ,&nbsp;Nanda Ramchandar ,&nbsp;Shelley M. Lawrence ,&nbsp;Aparajita Karmakar ,&nbsp;Ankit Sharma ,&nbsp;Mridu Sinha ,&nbsp;David T. Pride ,&nbsp;Peiting Kuo ,&nbsp;Khrissa Lecrone ,&nbsp;Megan Chiu ,&nbsp;Karen K. Mestan ,&nbsp;Eniko Sajti ,&nbsp;Michelle Vanderpool ,&nbsp;Sarah Lazar ,&nbsp;Melanie Crabtree ,&nbsp;Yordanos Tesfai ,&nbsp;Stephanie I. Fraley","doi":"10.1016/j.jmoldx.2024.01.013","DOIUrl":"10.1016/j.jmoldx.2024.01.013","url":null,"abstract":"<div><p>Fast and accurate diagnosis of bloodstream infection is necessary to inform treatment decisions for septic patients, who face hourly increases in mortality risk. Blood culture remains the gold standard test but typically requires approximately 15 hours to detect the presence of a pathogen. We, therefore, assessed the potential for universal digital high-resolution melt (U-dHRM) analysis to accomplish faster broad-based bacterial detection, load quantification, and species-level identification directly from whole blood. Analytical validation studies demonstrated strong agreement between U-dHRM load measurement and quantitative blood culture, indicating that U-dHRM detection is highly specific to intact organisms. In a pilot clinical study of 17 whole blood samples from pediatric patients undergoing simultaneous blood culture testing, U-dHRM achieved 100% concordance when compared with blood culture and 88% concordance when compared with clinical adjudication. Moreover, U-dHRM identified the causative pathogen to the species level in all cases where the organism was represented in the melt curve database. These results were achieved with a 1-mL sample input and sample-to-answer time of 6 hours. Overall, this pilot study suggests that U-dHRM may be a promising method to address the challenges of quickly and accurately diagnosing a bloodstream infection.</p></div>","PeriodicalId":50128,"journal":{"name":"Journal of Molecular Diagnostics","volume":"26 5","pages":"Pages 349-363"},"PeriodicalIF":4.1,"publicationDate":"2024-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139921427","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}