title: High-throughput gender determination using automated denaturant gel capillary electrophoresis.
abstract: Sex determination of anonymous samples is a requirement before analysis of DNA variation on X or Y chromosomes. Based on this, we designed a method for screening samples on different DNA capillary sequencing instruments with a sensitivity that is able to quantify sex chromosome abnormalities. The two different amelogenin alleles sited on the X and Y chromosomes were polymerase chain reaction amplified with the same set of primers and separated by denaturant capillary electrophoresis (DCE). Sex chromosome ratios could be reproducibly determined with a relative standard deviation of 8.7%, which is sufficient to distinguish a normal XY karyotype from an XYY karyotype associated with Klinefelter syndrome. Reconstruction experiments demonstrated sensitivity down to a simulated Y:X allelic ratio of 1:127 in all three instruments, enabling the prediction of sex chromosomal aneuploidies. When tested on anonymous pooled and single samples, DCE gave a good prediction of the male to female ratio in pools of 1000 blood donors. In conclusion, DCE is a simple and robust method for sex determination that can be readily performed on commercially available CE systems.
authors.affiliation: Department of Surgical Oncology, The Norwegian Radium Hospital, Oslo, Norway.
authors.forename: David C
authors.initials: DC
authors.lastname: Hinselwood
authors.forename: David J
authors.initials: DJ
authors.lastname: Warren
authors.forename: Per O
authors.initials: PO
authors.lastname: Ekstrøm
mesh_terms: D000483:Alleles; D053523:Amelogenin; D003746:Dental Enamel Proteins / Q000235:genetics*; D019075:Electrophoresis, Capillary / Q000379:methods; D005260:Female; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D012732:Sex Determination Analysis / Q000379:methods*
chemical_list: D053523:Amelogenin; D003746:Dental Enamel Proteins