Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
4
|
| pubmed:dateCreated |
1993-12-7
|
| pubmed:abstractText |
Lesch-Nyhan syndrome is an X-linked recessive disorder caused by molecular defects within the HPRT gene. Deletional forms of this syndrome, most of which are inherited, account for 15% of the cases. In addition, a large percentage of cases are due to de novo point mutations. We have used complementary fluorescence-based PCR assays to analyse disease-causing mutations in three unrelated families: (1) inheritance of dye-labelled PCR products of linked polymorphic loci mapping within and flanking the HPRT gene; (2) dye-labelled exon dosage analysis and (3) automated fluorescence-based DNA sequence analysis. Our results using fluorescent, dye-tagged PCR products show that inheritance of two polymorphic small tandem repeats, HPRTB [AGAT]n, mapping within intron 3 of the HPRT gene, and the CA-repeat at DXS294 can be used to establish linkage to the disease. In addition, we modified a previously described PCR protocol to use fluorescent dye-labelled oligoprimers and an ABI Gene Scanner in order to rapidly quantitate deletional forms of Lesch-Nyhan syndrome. Quantitative PCR analysis of individual exons followed by dosage analysis confirmed a deletion encompassing exon 9. A similar approach was used to confirm a previously described HPRT gene duplication involving exons 2 and 3. In this analysis, we co-amplified the HPRTB [AGAT]n and HUMARA [AGC]n repeats and confirmed increased exon dosage in carriers for the duplication. DNA sequence analysis remains the method of choice for delineating new disease-causing mutations, most of which are non-deletional forms of Lesch-Nyhan syndrome. We have also used a cycle-sequencing strategy employing dye-labelled dideoxy terminators and a laser-activated, fluorescence-emission DNA sequencer in order to define carrier status in 10 family members at risk for Lesch-Nyhan syndrome due to a splice donor mutation in intron 7. Our DNA sequence analyses corroborate small tandem repeat (STR) inheritance patterns in this family. Multiple fluorescence-based strategies should facilitate rapid diagnosis of the various Lesch-Nyhan disease-causing mutations.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
Aug
|
| pubmed:issn |
0890-8508
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:volume |
7
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
311-24
|
| pubmed:dateRevised |
2010-11-18
|
| pubmed:meshHeading |
pubmed-meshheading:8232348-Adolescent,
pubmed-meshheading:8232348-Adult,
pubmed-meshheading:8232348-Base Sequence,
pubmed-meshheading:8232348-DNA Primers,
pubmed-meshheading:8232348-Female,
pubmed-meshheading:8232348-Genetic Linkage,
pubmed-meshheading:8232348-Heterozygote,
pubmed-meshheading:8232348-Humans,
pubmed-meshheading:8232348-Hypoxanthine Phosphoribosyltransferase,
pubmed-meshheading:8232348-Introns,
pubmed-meshheading:8232348-Lesch-Nyhan Syndrome,
pubmed-meshheading:8232348-Male,
pubmed-meshheading:8232348-Molecular Sequence Data,
pubmed-meshheading:8232348-Multigene Family,
pubmed-meshheading:8232348-Pedigree,
pubmed-meshheading:8232348-Point Mutation,
pubmed-meshheading:8232348-Polymerase Chain Reaction,
pubmed-meshheading:8232348-Polymorphism, Genetic,
pubmed-meshheading:8232348-Repetitive Sequences, Nucleic Acid,
pubmed-meshheading:8232348-Sequence Analysis, DNA,
pubmed-meshheading:8232348-Sequence Deletion
|
| pubmed:year |
1993
|
| pubmed:articleTitle |
Fluorescent approaches to diagnosis of Lesch-Nyhan syndrome and quantitative analysis of carrier status.
|
| pubmed:affiliation |
Applied Biosystems, Inc., Foster City, CA.
|
| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|