title: Multiplex DNA deletion detection and exon sequencing of the hypoxanthine phosphoribosyltransferase gene in Lesch-Nyhan families.
abstract: The Lesch-Nyhan (LN) syndrome is a genetically lethal human neurological disease that results from mutations that inactivate the hypoxanthine phosphoribosyltransferase (HPRT) gene. The elucidation of the complete DNA sequence of the human HPRT gene locus has enabled the construction of multiple oligonucleotide primer sets for the simultaneous in vitro amplification of all nine HPRT exons. The multiplex polymerase chain reaction provides a facile assay for the detection of HPRT exon deletions and the reaction products can be analyzed by direct automated fluorescent DNA sequencing to identify subtle alterations in the gene. Alterations have been identified in the HPRT genes from 15 independent LN cases, and 10 LN family studies were performed. The sequencing method uses solid supports and is sufficiently simple and sensitive to be a favored approach for LN diagnosis. LN heterozygotes can be diagnosed without reference to the affected male. In addition, these procedures will be useful for somatic mutagenesis studies.
authors.affiliation: Institute for Molecular Genetics, Howard Hughes Medical Institute, Houston, Texas.
authors.forename: R A
authors.initials: RA
authors.lastname: Gibbs
authors.forename: P N
authors.initials: PN
authors.lastname: Nguyen
authors.forename: A
authors.initials: A
authors.lastname: Edwards
authors.forename: A B
authors.initials: AB
authors.lastname: Civitello
authors.forename: C T
authors.initials: CT
authors.lastname: Caskey
mesh_terms: D001483:Base Sequence; D002872:Chromosome Deletion; D004247:DNA / Q000235:genetics; D005091:Exons; D005260:Female; D005796:Genes; D006580:Genetic Carrier Screening; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics*; D007926:Lesch-Nyhan Syndrome / Q000235:genetics*; D008297:Male; D008969:Molecular Sequence Data; D010375:Pedigree; D016133:Polymerase Chain Reaction
chemical_list: D004247:DNA; D007041:Hypoxanthine Phosphoribosyltransferase