title: Convenient and quantitative determination of the frequency of a mutant allele using solid-phase minisequencing: application to aspartylglucosaminuria in Finland.
abstract: Aspartylglucosaminuria (AGU) is a recessively inherited lysosomal disease caused by inadequate aspartylglucosaminidase (AGA) activity. The disease is prevalent in the genetically isolated Finnish population. We have used a new method, solid-phase minisequencing, to determine the frequency of two missense mutations in the AGA gene in this population. In samples from 70% of the Finnish AGU families, we found that the two nucleotide changes were always associated, and they were identified in 98% of the AGU alleles analyzed. Thus, the high prevalence of AGU in the Finnish population is the consequence of a founder effect of one ancient mutation. The identification of asymptomatic carriers by the minisequencing test proved to be unequivocal. The method also allowed quantification of a mutated nucleotide sequence present in less than 1% of a sample. The frequency of AGU carriers in this population was 1/36 when estimated by quantifying the mutated AGU allele in a pooled leukocyte sample from 1350 normal Finnish individuals.
authors.affiliation: Laboratory of Molecular Genetics, National Public Health Institute, Helsinki, Finland.
authors.forename: A C
authors.initials: AC
authors.lastname: Syvänen
authors.forename: E
authors.initials: E
authors.lastname: Ikonen
authors.forename: T
authors.initials: T
authors.lastname: Manninen
authors.forename: M
authors.initials: M
authors.lastname: Bengtström
authors.forename: H
authors.initials: H
authors.lastname: Söderlund
authors.forename: P
authors.initials: P
authors.lastname: Aula
authors.forename: L
authors.initials: L
authors.lastname: Peltonen
mesh_terms: D000117:Acetylglucosamine / Q000031:analogs & derivatives* / Q000652:urine; D000483:Alleles; D001227:Aspartylglucosylaminase / Q000235:genetics*; D001483:Base Sequence; D004247:DNA / Q000097:blood / Q000235:genetics / Q000302:isolation & purification; D005387:Finland / Q000453:epidemiology; D005787:Gene Frequency; D006580:Genetic Carrier Screening; D005821:Genetic Techniques; D006801:Humans; D007962:Leukocytes / Q000502:physiology; D016464:Lysosomal Storage Diseases / Q000453:epidemiology / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction / Q000379:methods; D015995:Prevalence
chemical_list: D009838:Oligodeoxyribonucleotides; C000009:N-acetylglucosaminylasparagine; D004247:DNA; D001227:Aspartylglucosylaminase; D000117:Acetylglucosamine