[{"data":1,"prerenderedAt":-1},["ShallowReactive",2],{"detail-sidebar-cat-0-en-105":3,"doc-seo-331695-105":59,"doc-detail-331695-en":130},{"code":4,"msg":5,"data":6},0,"success",[7,13,18,23,28,33,38,43,48,51,55],{"id":8,"doc_module":4,"doc_module_name":9,"category_name":10,"show_sort_weight":11,"slug":12},1,"Document","Story & Novel",90,"story-novel",{"id":14,"doc_module":4,"doc_module_name":9,"category_name":15,"show_sort_weight":16,"slug":17},2,"Literature",80,"literature",{"id":19,"doc_module":4,"doc_module_name":9,"category_name":20,"show_sort_weight":21,"slug":22},4,"Exam",70,"exam",{"id":24,"doc_module":4,"doc_module_name":9,"category_name":25,"show_sort_weight":26,"slug":27},5,"Comic",60,"comic",{"id":29,"doc_module":4,"doc_module_name":9,"category_name":30,"show_sort_weight":31,"slug":32},6,"Technology",50,"technology",{"id":34,"doc_module":4,"doc_module_name":9,"category_name":35,"show_sort_weight":36,"slug":37},7,"Healthcare",40,"healthcare",{"id":39,"doc_module":4,"doc_module_name":9,"category_name":40,"show_sort_weight":41,"slug":42},8,"Research & Report",30,"research-report",{"id":44,"doc_module":4,"doc_module_name":9,"category_name":45,"show_sort_weight":46,"slug":47},9,"Religion & Spirituality",20,"religion-spirituality",{"id":46,"doc_module":4,"doc_module_name":9,"category_name":49,"show_sort_weight":46,"slug":50},"World Cup","world-cup",{"id":52,"doc_module":4,"doc_module_name":9,"category_name":53,"show_sort_weight":52,"slug":54},10,"Lifestyle","lifestyle",{"id":56,"doc_module":4,"doc_module_name":9,"category_name":57,"show_sort_weight":24,"slug":58},19,"General","general",{"code":4,"msg":60,"data":61},"ok",{"site_id":62,"language":63,"slug":64,"title":65,"keywords":66,"description":67,"schema_data":68,"social_meta":123,"head_meta":125,"extra_data":127,"updated_unix":129},105,"en","simultaneous-detection-of-eight-cancer-types-using-a-multiplex-droplet-digital-pcr-assay","Simultaneous detection of eight cancer types using a multiplex droplet digital PCR assay","","DNA methylation biomarkers are increasingly used for cancer detection because tumor-associated methylation changes can appear early in carcinogenesis and are consistent across many cases. Droplet digital PCR (ddPCR) is widely used for methylation detection, yet multi-cancer detection with multiple ddPCR targets had not been implemented. This study develops a multiplex ddPCR assay for eight frequent tumor types by selecting differentially methylated targets, validating three targets across tumor and normal adjacent samples, and combining two ddPCR assays to generate a shared read-out.",{"@graph":69,"@context":122},[70,84,105],{"@type":71,"itemListElement":72},"BreadcrumbList",[73,77,79,82],{"item":74,"name":75,"@type":76,"position":8},"https://docshare.wps.com","Home","ListItem",{"item":78,"name":9,"@type":76,"position":14},"https://docshare.wps.com/document/",{"item":80,"name":40,"@type":76,"position":81},"https://docshare.wps.com/document/research-report/",3,{"item":83,"name":65,"@type":76,"position":19},"https://docshare.wps.com/document/simultaneous-detection-of-eight-cancer-types-using-a-multiplex-droplet-digital-pcr-assay/331695/",{"url":83,"name":65,"@type":85,"image":86,"author":91,"headline":65,"publisher":94,"fileFormat":97,"inLanguage":63,"description":67,"dateModified":98,"datePublished":99,"encodingFormat":97,"isAccessibleForFree":100,"interactionStatistic":101},"DigitalDocument",{"url":87,"@type":88,"width":89,"height":90},"https://docshare.wps.com/thumbnails/simultaneous-detection-of-eight-cancer-types-using-a-multiplex-droplet-digital-pcr-assay/331695.png","ImageObject",300,407,{"name":92,"@type":93},"Aurelia","Person",{"url":74,"name":95,"@type":96},"DocShare","Organization","application/pdf","2026-10-28","2026-09-21",true,{"@type":102,"interactionType":103,"userInteractionCount":34},"InteractionCounter",{"@type":104},"ViewAction",{"@type":106,"mainEntity":107},"FAQPage",[108,114,118],{"name":109,"@type":110,"acceptedAnswer":111},"What is the main goal of the multiplex ddPCR assay described?","Question",{"text":112,"@type":113},"To enable simultaneous, highly accurate detection across eight frequent cancer types using methylation biomarkers measured by a multiplex droplet digital PCR approach.","Answer",{"name":115,"@type":110,"acceptedAnswer":116},"How were the methylation targets selected and validated?",{"text":117,"@type":113},"Targets were selected using prior analyses from The Cancer Genome Atlas (TCGA), then three targets were validated with ddPCR across tumor and normal adjacent fresh frozen samples.",{"name":119,"@type":110,"acceptedAnswer":120},"What performance does the assay achieve overall?",{"text":121,"@type":113},"The overall cross-validated area under the curve (cvAUC) is 0.948, with sensitivities ranging from 53.8% to 100% and specificities ranging from 80% to 100%.","https://schema.org",{"og:url":83,"og:type":124,"og:title":65,"og:site_name":95,"og:description":67},"article",{"robots":126,"canonical":83},"index,follow",{"doc_id":128,"site_id":62},331695,1790143701,{"code":4,"msg":5,"data":131},{"doc_id":128,"user_id":132,"nickname":92,"user_avatar":133,"doc_module":4,"category_id":39,"category_name":40,"doc_title":65,"doc_description":67,"doc_content":134,"file_id":135,"file_url":136,"file_type":137,"file_size":138,"view_count":34,"is_deleted":4,"is_public":8,"is_downloadable":8,"audit_status":8,"page_count":139,"language":140,"language_code":63,"site_id":62,"html_lang":63,"table_of_contents":141,"faqs":142,"seo_title":143,"seo_description":67,"update_tm":144,"read_time":36,"preview_page_count":4},1099514068365,"https://ap-avatar.wpscdn.com/avatar/10000253d8d9f28188e?x-image-process=image/resize,m_fixed,w_180,h_180&k=1794309523707217728","Simultaneous detection of eight cancer types using a multiplex droplet digital PCR assay  \nIsabelle Neefs 1,2 , Nele De Meulenaere 1,2, Thomas Vanpoucke 1,2, Janah Vandenhoeck 1,2, Dieter Peeters3, Marc Peeters2, Guy Van Camp 1,2 and Ken Op de Beeck 1,2   \n1 Center of Medical Genetics, University of Antwerp and Antwerp University Hospital, Edegem, Belgium  \n2 Center for Oncological Research, University of Antwerp and Antwerp University Hospital, Wilrijk, Belgium  \n3 Department of Pathology, Antwerp University Hospital, Edegem, Belgium  \nKeywords  \nbiomarkers; ddPCR; DNA methylation; multicancer detection; multiplexing  \nCorrespondence  \nK. Op de Beeck, Center of Medical Genetics, University of Antwerp and Antwerp University Hospital, Prins Boudewijnlaan 43, 2650 Edegem, Belgium Tel: +32 3 275 97 91  \nE-mail: ken.opdebeeck@uantwerpen. be  \n(Received 3 January 2024, revised 10 June 2024, accepted 23 July 2024, available online 6 September 2024)  \ndoi:10 . 1002/1878-0261 .13708  \nDNA methylation biomarkers have emerged as promising tools for cancer detection. Common methylation patterns across tumor types allow multi-cancer detection. Droplet digital PCR (ddPCR) has gained considerable attention for methylation detection. However, multi-cancer detection using multiple targets in ddPCR has never been performed before. Therefore, we developed a multiplex ddPCR assay for multi-cancer detection. Based on previous data analyses using The Cancer Genome Atlas (TCGA), we selected differentially methylated targets for eight frequent tumor types (lung, breast, colorectal, prostate, pancreatic, head and neck, liver, and esophageal cancer) . Three targets were validated using ddPCR in 103 tumor and 109 normal adjacent fresh frozen samples. Two distinct ddPCR assays were successfully developed. Output data from both assays is combined to obtain a read-out from the three targets together. Our overall ddPCR assay has a cross-validated area under the curve (cvAUC) of 0 .948. Performance between distinct cancer types varies, with sensitivities ranging from 53.8% to 100% and specificities ranging from 80% to 100% . Compared to previously published single-target parameters, we show that combining targets can drastically increase sensitivity and specificity, while lowering DNA input. In conclusion, we are the first to report a multi-cancer methylation ddPCRassay, which allows for highly accurate tumor predictions.  \n1. Introduction  \nCancer is one of the leading causes of death worldwide, with nearly 10 million deaths reported in 2020 . The most common cancers are breast (2.26 million new cases), lung (2.21 million new cases), colorectal (1.93 million new cases) and prostate (1 .41 million new cases) [1] . Many cancers can be cured when treated efficiently and, more importantly, detected early. However, patients are still  \nfrequently diagnosed in a late stage [2], despite efforts in screening programs and early detection methods. For example, 68% of lung cancer and 59% of colorectal cancer patients is diagnosed in stage IV [2], where 5-year survival rates are 7% and 14%, respectively. There are currently only screening programs in place for colorectal, breast, and cervical cancer in most western countries,  \nAbbreviations  \n%CV, intra-class coefficient of variability; cfDNA, cell-free DNA; cvAUC, cross-validated area under the curve; ddPCR, droplet digital PCR; EMX1, empty spiracles-like protein 1; HCC, hepatocellular carcinoma; LOB, limit of blank; LOD, limit of detection; NFQ-MGB, nonfluorescent quencher minor groove binder; NXPH1, neurexophilin 1; ROC, receiver-operator characteristic; TCGA, The Cancer Genome Atlas; TcP, tumor cell percentage; TOO, tissue of origin.  \n188 Molecular Oncology 19 (2025) 188–203 ª 2024 The Author(s) . Molecular Oncology published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.  \nThis is an open access article under the terms of the Creative Commons Attribution License, which permits use","cbCaipddJmQTMZQC","https://ap.wps.com/l/cbCaipddJmQTMZQC","pdf",1576061,16,"English","# Introduction\n## Rationale for early detection using DNA methylation\n## Limits of current screening and value of multi-cancer approaches","[{\"question\":\"What is the main goal of the multiplex ddPCR assay described?\",\"answer\":\"To enable simultaneous, highly accurate detection across eight frequent cancer types using methylation biomarkers measured by a multiplex droplet digital PCR approach.\"},{\"question\":\"How were the methylation targets selected and validated?\",\"answer\":\"Targets were selected using prior analyses from The Cancer Genome Atlas (TCGA), then three targets were validated with ddPCR across tumor and normal adjacent fresh frozen samples.\"},{\"question\":\"What performance does the assay achieve overall?\",\"answer\":\"The overall cross-validated area under the curve (cvAUC) is 0.948, with sensitivities ranging from 53.8% to 100% and specificities ranging from 80% to 100%.\"}]","Simultaneous detection of eight cancer types using a multiplex droplet digital PCR assay | PDF",1789994700]