{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,9,17]],"date-time":"2025-09-17T16:44:14Z","timestamp":1758127454298},"reference-count":20,"publisher":"Oxford University Press (OUP)","issue":"6","license":[{"start":{"date-parts":[[2008,6,1]],"date-time":"2008-06-01T00:00:00Z","timestamp":1212278400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2008,6,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Background: Huntington disease (HD) is a fatal autosomal dominant neurodegenerative disorder caused by an unstable expansion of the CAG trinucleotide repeat in exon 1 of the HTT (huntingtin) gene and typically has an adult onset. Molecular diagnosis and screening for HD currently involve separate amplification and detection steps.<\/jats:p>\n               <jats:p>Methods: We evaluated a novel, rapid microplate-based screening method for HD that combines the amplification and detection procedures in a single-step, closed-tube format. We carried out both the PCR for the HTT CAG-repeat region and the subsequent automated melting-curve analysis of the amplicon in the same wells on the plate. To establish cutoff melting temperatures (Tms) for each allelic class, we used a panel of reference DNA samples of known CAG-repeat sizes that represent a range of HTT alleles [normal (\u226426 repeats), intermediate (27\u201335 repeats), reduced penetrance expanded (36\u201339 repeats), and fully penetrant expanded (\u226540 repeats)]. We also measured well-to-well variation in Tm across the thermal block and validated cutoff Tms with DNA samples from 5 different populations. We also conducted a blinded validation analysis of clinical samples from an additional 40 HD-affected and 30 unaffected individuals.<\/jats:p>\n               <jats:p>Results: We observed a strong correlation between CAG-repeat size and amplicon Tm among the reference DNA samples. Use of the Tm cutoffs we established revealed that 5 samples from unaffected individuals had been misclassified as affected (1.1% false-positive rate). All samples from HD-affected and unaffected individuals were correctly identified in the blinded analysis.<\/jats:p>\n               <jats:p>Conclusions: This simple and scalable homogeneous assay may serve as a convenient, rapid, and accurate screen to detect the presence of pathologic expanded HD alleles in symptomatic patients.<\/jats:p>","DOI":"10.1373\/clinchem.2007.096503","type":"journal-article","created":{"date-parts":[[2008,4,11]],"date-time":"2008-04-11T01:09:15Z","timestamp":1207876155000},"page":"964-972","source":"Crossref","is-referenced-by-count":9,"title":["Single-Step Scalable-Throughput Molecular Screening for Huntington Disease"],"prefix":"10.1093","volume":"54","author":[{"given":"Clara R L","family":"Teo","sequence":"first","affiliation":[{"name":"Department of Pediatrics, Yong Loo Lin School of Medicine, National University of Singapore, Singapore"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wen","family":"Wang","sequence":"first","affiliation":[{"name":"Children\u2019s Medical Institute, National University Hospital, Singapore"},{"name":"Preimplantation Genetic Diagnosis Center, National University Hospital, Singapore"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hai","family":"Yang Law","sequence":"first","affiliation":[{"name":"Department of Pediatric Medicine, KK Women\u2019s and Children\u2019s Hospital, Singapore"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Caroline G","family":"Lee","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Samuel S","family":"Chong","sequence":"first","affiliation":[{"name":"Department of Pediatrics, Yong Loo Lin School of Medicine, National University of Singapore, Singapore"},{"name":"Children\u2019s Medical Institute, National University Hospital, Singapore"},{"name":"Preimplantation Genetic Diagnosis Center, National University Hospital, Singapore"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2008,6,1]]},"reference":[{"key":"2020022800141717700_R1","doi-asserted-by":"crossref","first-page":"971","DOI":"10.1016\/0092-8674(93)90585-E","volume":"72","year":"1993","journal-title":"Cell"},{"key":"2020022800141717700_R2","doi-asserted-by":"crossref","first-page":"398","DOI":"10.1038\/ng0893-398","volume":"4","year":"1993","journal-title":"Nat Genet"},{"key":"2020022800141717700_R3","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1136\/jmg.31.5.377","volume":"31","year":"1994","journal-title":"J Med Genet"},{"key":"2020022800141717700_R4","doi-asserted-by":"crossref","first-page":"1243","DOI":"10.1086\/301846","volume":"62","year":"1998","journal-title":"Am J Hum 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