title: A rare mutation in the primer binding region of the amelogenin gene can interfere with gender identification.
abstract: PCR amplification of part of the X-Y homologous amelogenin gene with a single primer pair has been used as a sex identification test because it generates different length products from the X and Y chromosomes. Using a commercially available kit that contains amelogenin primers, we report a single phenotypically normal Caucasian male out of 327 males tested to date that failed to show an X chromosome-specific PCR product. Using alternative amelogenin primers external to but encompassing the initial amplicon, an X chromosome-specific product was seen. Sequence analysis of this X-specific PCR product revealed a C to G mutation at the most 3' base of the initial reverse amelogenin PCR primer. An alternative reverse PCR primer with this most 3' base deleted showed X- and Y-specific products from the case study male. Rare mutations that result in a failure to amplify sex chromosome-specific products can result in incorrect gender identification.
authors.affiliation: Department of Clinical Pathology/L30, Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland, OH 44195, USA.
authors.forename: Bonnie
authors.initials: B
authors.lastname: Shadrach
authors.forename: Mairead
authors.initials: M
authors.lastname: Commane
authors.forename: Carol
authors.initials: C
authors.lastname: Hren
authors.forename: Ilka
authors.initials: I
authors.lastname: Warshawsky
mesh_terms: D000483:Alleles; D053523:Amelogenin; D041321:Chromosomes, Human, X; D017931:DNA Primers / Q000737:chemistry; D003746:Dental Enamel Proteins / Q000235:genetics*; D005260:Female; D006801:Humans; D008297:Male; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D011485:Protein Binding; D017422:Sequence Analysis, DNA / Q000379:methods; D012732:Sex Determination Analysis / Q000379:methods*
chemical_list: D053523:Amelogenin; D017931:DNA Primers; D003746:Dental Enamel Proteins