title: Prenatal diagnosis of trisomy 21, 18 and 13 by quantitative pyrosequencing of segmental duplications.
abstract: Chromosomal aberration mostly occurs in chromosomes 21, 18 and 13, with an incidence approximately 1 out of 160 live births in humans, therefore making prenatal diagnosis necessary in clinics. Current methods have drawbacks such as time consuming, high cost, complicated operations and low sensitivity. In this paper, a novel method for rapid and accurate prenatal diagnosis of aneuploidy is proposed based on pyrosequencing, which quantitatively detects the peak height ratio (PHR) of different bases of segmental duplication. A direct polymerase chain reaction (PCR) approach was undertaken, where a small volume of amniotic fluid was used as the starting material without DNA extraction. Single-stranded DNA was prepared from PCR products and subsequently analyzed using pyrosequencing. The PHR between target and reference chromosome of 2.2 for euploid and 3:2 for a trisomy fetus were used as reference. The reference intervals and z scores were calculated for discrimination of aneuploidy. A total of 132 samples were collected, within trisomy 21 (n = 11), trisomy 18 (n = 3), trisomy 13 (n = 2), and unaffected controls (n = 116). A set of six segmental duplications were chosen for analysis. This method had consistent results with karyotyping analysis, a correct diagnosis with 100% sensitivity and 99.9% specificity.
authors.affiliation: Key Laboratory of Drug Quality Control and Pharmacovigilance of Ministry of Education, China Pharmaceutical University, Nanjing, China.
authors.forename: H
authors.initials: H
authors.lastname: Tong
authors.affiliation: Key Laboratory of Drug Quality Control and Pharmacovigilance of Ministry of Education, China Pharmaceutical University, Nanjing, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Jin
authors.affiliation: Key Laboratory of Drug Quality Control and Pharmacovigilance of Ministry of Education, China Pharmaceutical University, Nanjing, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Department of pharmacology, Jinling Hospital, Medical School of Nanjing University, Nanjing, China.
authors.forename: B
authors.initials: B
authors.lastname: Zou
authors.affiliation: Department of pharmacology, Jinling Hospital, Medical School of Nanjing University, Nanjing, China.
authors.forename: H
authors.initials: H
authors.lastname: Ye
authors.affiliation: Department of pharmacology, Jinling Hospital, Medical School of Nanjing University, Nanjing, China.
authors.forename: H
authors.initials: H
authors.lastname: Wu
authors.affiliation: School of Biological Sciences, Victoria University of Wellington, Wellington, New Zealand.
authors.forename: S
authors.initials: S
authors.lastname: Kumar
authors.affiliation: School of Biological Sciences, Victoria University of Wellington, Wellington, New Zealand.
authors.forename: J L
authors.initials: JL
authors.lastname: Pitman
authors.affiliation: Key Laboratory of Drug Quality Control and Pharmacovigilance of Ministry of Education, China Pharmaceutical University, Nanjing, China. ghzhou@nju.edu.cn.|Department of pharmacology, Jinling Hospital, Medical School of Nanjing University, Nanjing, China. ghzhou@nju.edu.cn.
authors.forename: G
authors.initials: G
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Drug Quality Control and Pharmacovigilance of Ministry of Education, China Pharmaceutical University, Nanjing, China. songqinxin@cpu.edu.cn.|Department of pharmacology, Jinling Hospital, Medical School of Nanjing University, Nanjing, China. songqinxin@cpu.edu.cn.
authors.forename: Q
authors.initials: Q
authors.lastname: Song
mesh_terms: D002869:Chromosome Aberrations; D025063:Chromosome Disorders / Q000175:diagnosis* / Q000235:genetics; D002882:Chromosomes, Human, Pair 13 / Q000235:genetics; D002887:Chromosomes, Human, Pair 18 / Q000235:genetics; D002891:Chromosomes, Human, Pair 21 / Q000235:genetics; D004277:DNA, Single-Stranded / Q000235:genetics; D004314:Down Syndrome / Q000175:diagnosis* / Q000235:genetics / Q000503:physiopathology; D005260:Female; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D007621:Karyotyping; D011247:Pregnancy; D011296:Prenatal Diagnosis; D056916:Segmental Duplications, Genomic / Q000235:genetics; D014314:Trisomy / Q000175:diagnosis* / Q000235:genetics / Q000503:physiopathology; D000073839:Trisomy 13 Syndrome; D000073842:Trisomy 18 Syndrome
chemical_list: D004277:DNA, Single-Stranded
keywords: amniotic fluid amplification; chromosomal aneuploidy; prenatal diagnosis; pyrosequencing; segmental duplication