title: Rapid diagnosis of aneuploidy using segmental duplication quantitative fluorescent PCR.
abstract: The aim of this study was use a simple and rapid procedure, called segmental duplication quantitative fluorescent polymerase chain reaction (SD-QF-PCR), for the prenatal diagnosis of fetal chromosomal aneuploidies. This method is based on the co-amplification of segmental duplications located on two different chromosomes using a single pair of fluorescent primers. The PCR products of different sizes were subsequently analyzed through capillary electrophoresis, and the aneuploidies were determined based on the relative dosage between the two chromosomes. Each primer set, containing five pairs of primers, was designed to simultaneously detect aneuploidies located on chromosomes 21, 18, 13, X and Y in a single reaction. We applied these two primer sets to DNA samples isolated from individuals with trisomy 21 (n = 36); trisomy 18 (n = 6); trisomy 13 (n = 4); 45, X (n = 5); 47, XXX (n = 3); 48, XXYY (n = 2); and unaffected controls (n = 40). We evaluated the performance of this method using the karyotyping results. A correct and unambiguous diagnosis with 100% sensitivity and 100% specificity, was achieved for clinical samples examined. Thus, the present study demonstrates that SD-QF-PCR is a robust, rapid and sensitive method for the diagnosis of common aneuploidies, and these analyses can be performed in less than 4 hours for a single sample, providing a competitive alternative for routine use.
authors.affiliation: Prenatal Diagnosis Center, the First Affiliated Hospital of Zhengzhou University, Henan, China.
authors.forename: Xiangdong
authors.initials: X
authors.lastname: Kong
authors.affiliation: Department of Genetic Laboratory, Lin Yi People's Hospital, Shandong, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Li
authors.affiliation: Laboratory of Medical Genetics, Qinzhou Maternal and Child Health Care Hospital, Guangxi, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Sun
authors.affiliation: Laboratory of Medical Genetics, Qinzhou Maternal and Child Health Care Hospital, Guangxi, China.
authors.forename: Kepeng
authors.initials: K
authors.lastname: Fu
authors.affiliation: Laboratory of Medical Genetics, Qinzhou Maternal and Child Health Care Hospital, Guangxi, China.
authors.forename: Ju
authors.initials: J
authors.lastname: Long
authors.affiliation: Laboratory of Medical Genetics, Qinzhou Maternal and Child Health Care Hospital, Guangxi, China.
authors.forename: Xunjin
authors.initials: X
authors.lastname: Weng
authors.affiliation: Laboratory of Medical Genetics, Qinzhou Maternal and Child Health Care Hospital, Guangxi, China.
authors.forename: Xuehe
authors.initials: X
authors.lastname: Ye
authors.affiliation: Laboratory of Medical Genetics, Qinzhou Maternal and Child Health Care Hospital, Guangxi, China.
authors.forename: Xinxiong
authors.initials: X
authors.lastname: Liu
authors.affiliation: Genetics Laboratory, Hubei Maternal and Child Health Hospital, Hubei, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Wang
authors.affiliation: Laboratory of Medical Genetics, Qinzhou Maternal and Child Health Care Hospital, Guangxi, China.
authors.forename: Shanhuo
authors.initials: S
authors.lastname: Yan
authors.affiliation: Laboratory of Medical Genetics, Qinzhou Maternal and Child Health Care Hospital, Guangxi, China.
authors.forename: Haiming
authors.initials: H
authors.lastname: Ye
authors.affiliation: Laboratory of Medical Genetics, Qinzhou Maternal and Child Health Care Hospital, Guangxi, China.
authors.forename: Zuqian
authors.initials: Z
authors.lastname: Fan
mesh_terms: D000782:Aneuploidy; D001331:Automation; D025063:Chromosome Disorders / Q000175:diagnosis*; D002874:Chromosome Mapping; D017931:DNA Primers / Q000737:chemistry / Q000235:genetics; D019075:Electrophoresis, Capillary; D005260:Female; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D007621:Karyotyping; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D011296:Prenatal Diagnosis / Q000379:methods*; D015203:Reproducibility of Results; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D014314:Trisomy / Q000235:genetics*
chemical_list: D017931:DNA Primers; D005456:Fluorescent Dyes