Multiplex DNA deletion detection and exon sequencing of the hypoxanthine phosphoribosyltransferase gene in Lesch-Nyhan families.

TitleMultiplex DNA deletion detection and exon sequencing of the hypoxanthine phosphoribosyltransferase gene in Lesch-Nyhan families.
Publication TypeJournal Article
Year of Publication1990
AuthorsGibbs, RA, Nguyen, PN, Edwards, A, Civitello, AB, Caskey, CT
JournalGenomics
Volume7
Issue2
Pagination235-44
Date Published1990 Jun
ISSN0888-7543
KeywordsBase Sequence, Chromosome Deletion, DNA, Exons, Female, Genes, Genetic Carrier Screening, Humans, Hypoxanthine Phosphoribosyltransferase, Lesch-Nyhan Syndrome, Male, Molecular Sequence Data, Pedigree, Polymerase Chain Reaction
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.

Alternate JournalGenomics
PubMed ID2347587
Grant ListDK31428 / DK / NIDDK NIH HHS / United States