[{"data":1,"prerenderedAt":-1},["ShallowReactive",2],{"detail-sidebar-cat-0-en-105":3,"doc-seo-452748-105":59,"doc-detail-452748-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","biochemical-testing-promotes-interpretation-of-variants-of-uncertain-significance-in-prenatal-genetic-disease-testing-in-four-organic-acidurias","Biochemical Testing Promotes Interpretation of Variants of Uncertain Significance in Prenatal Genetic Disease Testing in Four Organic Acidurias","","Advances in next-generation sequencing (NGS) have transformed diagnosis of genetic disorders, including newborn and prenatal screening, but the interpretation of variants of uncertain significance (VUS) continues to lag. VUS are genetic alterations with insufficient evidence for reliable clinical classification and are often not immediately actionable, with misclassification carrying serious consequences. This open-access review synthesizes evidence on amniotic fluid metabolite analysis for prenatal diagnosis in four organic acidurias, outlining how biochemical testing supports VUS interpretation and strengthens clinical decision-making.",{"@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":35,"@type":76,"position":81},"https://docshare.wps.com/document/healthcare/",3,{"item":83,"name":65,"@type":76,"position":19},"https://docshare.wps.com/document/biochemical-testing-promotes-interpretation-of-variants-of-uncertain-significance-in-prenatal-genetic-disease-testing-in-four-organic-acidurias/452748/",{"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/biochemical-testing-promotes-interpretation-of-variants-of-uncertain-significance-in-prenatal-genetic-disease-testing-in-four-organic-acidurias/452748.png","ImageObject",300,407,{"name":92,"@type":93},"acoobca","Person",{"url":74,"name":95,"@type":96},"DocShare","Organization","application/pdf","2026-11-07","2026-09-30",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},"Why do variants of uncertain significance (VUS) remain challenging in prenatal genetic testing?","Question",{"text":112,"@type":113},"VUS lack sufficient evidence for clear pathogenicity or benignity and therefore are often not immediately actionable. Their classification can also change over time as new evidence emerges.","Answer",{"name":115,"@type":110,"acceptedAnswer":116},"How does biochemical testing help interpret VUS?",{"text":117,"@type":113},"Functional studies and clinically available biochemical findings can provide evidence about variant effects on gene/protein function or fetal metabolic phenotype. In the review, amniotic fluid metabolite analysis is presented as a rapid adjunct that may support VUS reclassification.",{"name":119,"@type":110,"acceptedAnswer":120},"What is the rationale for using amniotic fluid metabolite analysis in prenatal diagnosis?",{"text":121,"@type":113},"Mass spectrometry can assess fetal metabolic status using minimal sample volumes and generate results within days. Abnormal metabolite patterns in amniotic fluid provide phenotypic support for inherited metabolic disorder diagnoses and can strengthen evidence categories such as PP4.","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},452748,1791296305,{"code":4,"msg":5,"data":131},{"doc_id":128,"user_id":132,"nickname":92,"user_avatar":133,"doc_module":4,"category_id":34,"category_name":35,"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":34,"language":139,"language_code":63,"site_id":62,"html_lang":63,"table_of_contents":140,"faqs":141,"seo_title":142,"seo_description":67,"update_tm":143,"read_time":144,"preview_page_count":4},3985747870733,"https://ap-avatar.wpscdn.com/davatar_a8503ba1806abce46bf441b54a3ca4cd","Clinical Genetics  \nREVIEW  OPEN ACCESS   \nBiochemical Testing Promotes Interpretation of Variants of Uncertain Significance in Prenatal Genetic Disease Testing in Four Organic Acidurias  \nKaili Yin | Qingwei Qi   \nDepartment of Obstetrics and Gynecology, National Clinical Research Center for Obstetric & Gynecologic Diseases, Peking Union Medical College Hospital,  \nChinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China Correspondence: Qingwei Qi ([qiqingwei@163.com](qiqingwei@163.com))  \nReceived: 11 June 2025 | Revised: 25 September 2025 | Accepted: 26 September 2025  \nFunding: This work was supported by the National Key Clinical Specialty Construction Project (U114000) .  \nKeywords: biochemical testing | prenatal diagnosis | variants of uncertain significance  \nABSTRACT  \nAdvances in next-generation sequencing (NGS) have revolutionized clinical and translational medicine. NGS is increasingly used in the diagnosis of genetic diseases, including newborn screening, prenatal screening and diagnosis, and preimplantation screening. While advancements in sequencing and bioinformatics have facilitated the discovery of novel genes and diseases, the identification of variants continues to outpace our ability to classify them accurately. Variants of uncertain significance (VUS), defined as genetic alterations with insufficient evidence for clinical interpretation, present significant diagnostic challenges. Clinically available functional studies, such as biochemical testing, offer valuable insights for interpreting VUS. This review synthesizes recent literature on amniotic fluid metabolite analysis for prenatal diagnosis and VUS interpretation in four organic acidurias. It aims to guide the application of biochemical testing for inherited metabolic disorders and enhance clinical decisionmaking regarding VUS classification.  \n1 | Introduction  \nThe implementation of next-generation sequencing (NGS) technologies, particularly whole exome sequencing (WES) and whole genome sequencing, has revolutionized the diagnosis of genetic disorders over the past decade. Despite improved diagnostic rates, approximately 60%–70% of patients still lack a definitive genetic diagnosis [1, 2] . This gap is partly due to the complex interpretation of the numerous variants of uncertain significance (VUS), a category within the five-tier classification system of the American College of Medical Genetics and Genomics (ACMG). VUS are genetic variants lacking sufficient evidence for definitive clinical interpretation. VUS typically fall into two categories: (1) variants with insufficient evidence for pathogenicity/benignity due to limited data; or (2) variants with conflicting evidence supporting both pathogenicity and  \nbenignity. The classification of VUS may change over time. However, from a clinical perspective, the identification of VUS presents a significant challenge, as VUS are not immediately actionable. Furthermore, novel variants unreported in the literature are challenging to interpret. Misclassification can lead to serious consequences for patients and their families [3–6] .  \nAccording to the Sequence Variant Interpretation Criteria and Guidelines of the ACMG, VUS should not be used to guide clinical decision-making [7]. ACMG and the Association for Molecular Pathology (AMP) jointly developed standards and guidelines for the assessment of evidence to increase consistency and transparency in clinical variant interpretation [7]. One type of evidence defined in this guideline was the effect of a variant on gene/protein function determined by a “well-established” functional assay, providing strong support of a pathogenic or benign impact (codes  \nThis is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.  ","cbCaiud4kL927IcA","https://ap.wps.com/l/cbCaiud4kL927IcA","pdf",261201,"English","# 1 | Introduction\n## Next-generation sequencing and the diagnostic gap for VUS\n## ACMG/AMP guidelines and functional evidence\n## Limits of in vitro functional assays in prenatal timelines\n## Amniotic fluid metabolite analysis and PP4/biochemical evidence\n## Biochemical testing targets across organic acidurias","[{\"question\":\"Why do variants of uncertain significance (VUS) remain challenging in prenatal genetic testing?\",\"answer\":\"VUS lack sufficient evidence for clear pathogenicity or benignity and therefore are often not immediately actionable. Their classification can also change over time as new evidence emerges.\"},{\"question\":\"How does biochemical testing help interpret VUS?\",\"answer\":\"Functional studies and clinically available biochemical findings can provide evidence about variant effects on gene/protein function or fetal metabolic phenotype. In the review, amniotic fluid metabolite analysis is presented as a rapid adjunct that may support VUS reclassification.\"},{\"question\":\"What is the rationale for using amniotic fluid metabolite analysis in prenatal diagnosis?\",\"answer\":\"Mass spectrometry can assess fetal metabolic status using minimal sample volumes and generate results within days. Abnormal metabolite patterns in amniotic fluid provide phenotypic support for inherited metabolic disorder diagnoses and can strengthen evidence categories such as PP4.\"}]","Biochemical Testing Promotes Interpretation of Variants of Uncertain Significance in Prenatal Genetic Disease Testing in Four Organic Acidurias | PDF",1790737650,18]