title: Rapid diagnosis of aneuploidy by high-resolution melting analysis of segmental duplications.
abstract: BACKGROUND
  Several molecular methods, such as quantitative fluorescence PCR and multiplex ligation-dependent probe amplification, currently serve as important adjuncts to traditional karyotyping for the diagnosis of aneuploidy; however, the performance or throughput limitations of these methods hinder their use for routine prenatal diagnosis and population-based postnatal screening. We developed a novel approach, called "high-resolution melting analysis of segmental duplications," to detect common aneuploidies.
  
  
  METHODS
  In this method, similar sequences located on different chromosomes are amplified simultaneously with a single primer set; the PCR products are then analyzed by high-resolution melting. Aneuploidy-associated dosage abnormalities produce different ratios of similar amplicons, which produce melting curves that are detectably different from those of samples from unaffected individuals. We applied this method to DNA samples isolated from individuals with trisomy 21 (n = 48), trisomy 18 (n = 10), trisomy 13 (n = 3), 45,X (n = 8), and 47,XXY (n = 14), and from unaffected controls (n = 48).
  
  
  RESULTS
  As judged by the karyotyping results, our method attained 100% diagnostic sensitivity and 99.6% diagnostic specificity. Moreover, our method was able to detect a change in chromosome dosage as low as 1.05-fold.
  
  
  CONCLUSIONS
  This novel method clearly differentiates samples of patients with common aneuploidies from those of unaffected controls, while markedly simplifying the assays and reducing time and costs. The assay has sufficient throughput to meet the demands of large-scale testing, such as population-based postnatal screening, and is thus suitable for routine use.
authors.affiliation: Molecular Diagnostics Laboratory, Department of Medical Genetics, Prenatal Diagnosis Center of Xiamen, Maternal and Child Health Hospital, Xiamen, Fujian, China.
authors.forename: Qiwei
authors.initials: Q
authors.lastname: Guo
authors.forename: Li
authors.initials: L
authors.lastname: Xiao
authors.forename: Yulin
authors.initials: Y
authors.lastname: Zhou
mesh_terms: D000782:Aneuploidy; D025063:Chromosome Disorders / Q000175:diagnosis / Q000235:genetics; D002882:Chromosomes, Human, Pair 13 / Q000235:genetics; D002887:Chromosomes, Human, Pair 18 / Q000235:genetics; D004314:Down Syndrome / Q000175:diagnosis / Q000235:genetics; D005260:Female; D006801:Humans; D007621:Karyotyping; D008297:Male; D016133:Polymerase Chain Reaction; D011296:Prenatal Diagnosis / Q000379:methods; D056916:Segmental Duplications, Genomic; D012680:Sensitivity and Specificity; D025064:Sex Chromosome Disorders / Q000175:diagnosis / Q000235:genetics; D014314:Trisomy / Q000175:diagnosis / Q000235:genetics; D000073839:Trisomy 13 Syndrome; D014424:Turner Syndrome / Q000175:diagnosis / Q000235:genetics; D014997:XYY Karyotype / Q000175:diagnosis / Q000235:genetics