Document Detail

Improved polymerase chain reaction conditions for quick diagnostics of Huntington disease.
MedLine Citation:
PMID:  9438070     Owner:  NLM     Status:  MEDLINE    
Huntington disease (HD) belongs to a growing list of neurodegenerative disorders (fragile X syndrome [6], myotonic dystrophy [1], spino-bulbar muscular atrophy [2] etc.) characterized by unstable expanded trinucleotide repeats (so-called 'dynamic mutations'). The dynamic mutation causing HD represents the expansion of CAG triplets in the first exon of a gene IT15 (chromosome 4) coding for huntington. This trinucleotide stretch is varying in the range of 11-34 in normal chromosomes and 39-121 in HD chromosomes. The most direct diagnostic approach is to amplify the proximal region of IT15 gene (from patients genomic DNA) by polymerase chain reaction (PCR) and estimate the number of CAG triplets. All protocols published to date are difficult to reproduce because amplification is inefficient giving additional non-specific products. The strategy of our experiment is shown in Fig. 1. We designed one new primer, primer No. 2 (another primer was primer No. 1) and novel PCR conditions. Primer No. 2 is located closer to CAG triplets and its extension is not including the GC rich region. PCR amplified products, using primer Nos. 1 and 2, thus do not include the GC rich region and, therefore, are much more efficiently amplified (compared to the products of amplification with primer Nos. 1 and 3).
B Culjković; S Ruzdijić; L Rakić; S Romac
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Brain research. Brain research protocols     Volume:  2     ISSN:  1385-299X     ISO Abbreviation:  Brain Res. Brain Res. Protoc.     Publication Date:  1997 Dec 
Date Detail:
Created Date:  1998-02-26     Completed Date:  1998-02-26     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9716650     Medline TA:  Brain Res Brain Res Protoc     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  44-6     Citation Subset:  IM    
Institute for Biological Research, Belgrade, Yugoslavia.
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MeSH Terms
Chromosome Mapping
Chromosomes, Human, Pair 4
DNA Primers
Huntington Disease / blood,  diagnosis*,  genetics
Polymerase Chain Reaction / methods*
Trinucleotide Repeats
Reg. No./Substance:
0/DNA Primers

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