title: Rapid and simultaneous detection of common aneuploidies by quadruplex real-time polymerase chain reaction combined with melting curve analysis.
abstract: BACKGROUND
  During the prenatal period, the number variation of chromosomes 13, 18, 21, X and Y accounts for more than 80% of the clinically significant chromosomal abnormalities diagnosed. Rapid tests for prenatal diagnosis of these abnormalities can improve pregnancy management and alleviate parental anxiety. Here, we present a molecular alternative method for detecting common aneuploidies.
  
  
  METHODS
  This method is based on co-amplification of segmental duplications located on two different chromosomes using a single pair of primers. Segmental duplications have a high degree of sequence identity, but have single-nucleotide differences in some regions. These sequence differences can be quantified using melting curve analysis of dual-labeled probes to estimate the relative dosages of different chromosomes. We designed two quadruplex real-time PCR assays to detect aneuploidies of chromosomes 13, 18, 21, X and Y.
  
  
  RESULTS
  We examined 75 aneuploid DNA samples and 56 unaffected DNA control samples using these two assays and correctly identified all samples. Four cases of unbalanced translocation were also accurately detected. The observed averaged ratio for each chromosomal disorder was similar to the theoretically expected value.
  
  
  CONCLUSIONS
  Our real-time assay is a robust, rapid, and easy to conduct technique for prenatal diagnosis of common aneuploidies, representing a competitive alternative for use in diagnostic laboratories.
authors.affiliation: Prenatal Diagnosis Center, Dongguan Maternal and Child Health Hospital, Dongguan, Guangdong, People's Republic of China.
authors.forename: Jiwu
authors.initials: J
authors.lastname: Lou
authors.affiliation: Prenatal Diagnosis Center, Dongguan Maternal and Child Health Hospital, Dongguan, Guangdong, People's Republic of China.
authors.forename: Manna
authors.initials: M
authors.lastname: Sun
authors.affiliation: Prenatal Diagnosis Center, Dongguan Maternal and Child Health Hospital, Dongguan, Guangdong, People's Republic of China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Prenatal Diagnosis Center, Dongguan Maternal and Child Health Hospital, Dongguan, Guangdong, People's Republic of China.
authors.forename: Zhisong
authors.initials: Z
authors.lastname: Ji
authors.affiliation: Reproductive medicine Center, Guangdong Maternal and Child Health Hospital, Guangzhou, People's Republic of China.
authors.forename: Fenghua
authors.initials: F
authors.lastname: Liu
authors.affiliation: Prenatal Diagnostic Center, Guangzhou Women & Children Medical Center affiliated to Guangzhou Medical University, Guangzhou, Guangdong, People's Republic of China.
authors.forename: Dongzhi
authors.initials: D
authors.lastname: Li
authors.affiliation: Prenatal Diagnosis Center, Dongguan Maternal and Child Health Hospital, Dongguan, Guangdong, People's Republic of China.
authors.forename: Wanfang
authors.initials: W
authors.lastname: Xu
authors.affiliation: Prenatal Diagnosis Center, Dongguan Maternal and Child Health Hospital, Dongguan, Guangdong, People's Republic of China.
authors.forename: Yangyang
authors.initials: Y
authors.lastname: Lin
authors.affiliation: Prenatal Diagnosis Center, Dongguan Maternal and Child Health Hospital, Dongguan, Guangdong, People's Republic of China.
authors.forename: Yanhui
authors.initials: Y
authors.lastname: Liu
mesh_terms: D000782:Aneuploidy; D016022:Case-Control Studies; D025063:Chromosome Disorders / Q000175:diagnosis* / Q000235:genetics; D002882:Chromosomes, Human, Pair 13; D002887:Chromosomes, Human, Pair 18; D002891:Chromosomes, Human, Pair 21; D041321:Chromosomes, Human, X; D041322:Chromosomes, Human, Y; D004247:DNA / Q000032:analysis; D005260:Female; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D007621:Karyotyping; D008297:Male; D009711:Nucleotides; D011247:Pregnancy; D060888:Real-Time Polymerase Chain Reaction; D014314:Trisomy
chemical_list: D005456:Fluorescent Dyes; D009711:Nucleotides; D004247:DNA